Bulletin of Environment, Pharmacology and Life Sciences

Bull. Env. Pharmacol. Life Sci., Vol 9[[2] January 2020 : 164-177
©2020 Academy for Environment and Life Sciences, India
Online ISSN 2277-1808
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CODEN: BEPLAD
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REVIEW ARTICLE OPEN ACCESS

Ubiquity, Significance and the Molecular Techniques used for

deciphering Actinomycetes
Vishal Mathur, Indrani Jadhav*, Roshan Vasniwal, Divya Shrivastava and Kapilesh Jadhav
School of Life Science, Jaipur National University (Rajasthan), Jaipur 302017
Email: [email protected]

ABSTRACT
Actinomycetes are widely ly distributed in soil, water, air and in plant remains. Actinomycetes are present in extreme type
of habitats, Alkalophilic actinomycetes are present in alkaline soils, Cryobacterium psychrophilum with optimum growth
temperature at 9-12oC, acidophilic actinomycetes isolated from acidic forest and Psychrophilic actinomycetes colonies
were isolated from ice point region and some of the rare marine actinomycetes Salinispora require seawater for their
growth. A large number of the rare genera of actinomycetes
actinomycetes have been not explored yet with vast biotechnological and
industrial potential. With the assistance of molecular approaches and ongoing advances in genomics and sequencing
technologies, culture-independent
independent molecular techniques have started another periodperiod of actinomycetes environment. The
modern molecular technologies provide new source of chemical diversity with novel actinomycetes. The Streptomyces
family gives us the vital class of antibiotics that we are utilizing today, The Frankia family, works in n non leguminous
plants as nitrogen fixing organism, some Actinomycetes are likewise utilized in plant development (help to deliver plant
development hormone Indole-3-acidic).
acidic). Certain enzymes from actinomycetes example, amylase, lipase, and cellulases
play a vital part in textile, food, fermentation, agriculture as well as in paper industry. Actinomycetes are also
responsible in degrading pesticides, degrading hydrocarbons have major source of bioremediation and are also utilized
in biocorrosion.
Keywords: Alkalophilic
lkalophilic actinomycetes, Silinispora, cellulases, Hydrocarbons, Bioremediation.

Received 23.07.2019 Revised 15.11.2019 Accepted 04.12.2019

INTRODUCTION
The actinomycetes are gram-positive
positive bacteria with high G+C percentage that form branching pattern and
filamentous in nature. These bacteria closely resemble fungi in overall morphology ..Therefore, they are
also known as ray fungi. They are characterized by a complex life cycle belonging to the phylum
Actinobacteria. Actinobacteria occur in coccoid (Micrococcus),
( ), bar coccoid (e.g., Arthrobacter) to
partitioning hyphal shapes morphologically (e.g., Nocardia spp.) or forever and exceedingly isolated
spread mycelium (e.g., Streptomyces spp.)
spp It has been estimated that one-thirdthird of the thousands of
naturally occurring antibiotics have been produced from actinomycetes [1].. The increasingly antibiotic
resistant occur in the bacteria, especially occur in the Multidrug Resistant Microbes (MDRM) and
therefore need to look for new antibiotics [2].
Actinomycetes play a great role in the development of large
large number of bioactive compounds, which
after isolation, processing and characterization have been transformed into drugs for treating different
diseases in both plants and animals keeping this reality in view, actinomycetes are considered to be the
effective
ective sources for the generation of various antibiotics and other biologically active compounds. Each
strain of actinomycetes has the innate capability of producing approximately around 10 10-20 secondary
metabolites. Actinomycetes are extraordinary producers
producers of antibiotics and among actinomycetes the
major role is played by Streptomyces, which alone represent an outstanding 80% of the natural products
produced by actinomycetes [3]. Streptomyces have been a source to different analytics; including anti anti-
bacterial, anti-fungal and anti-cancer
cancer drugs [4]. Streptomyces accompanied by actinomycetes keep
maintaining the pace as a source of novel metabolites exhibiting various biological activities such as anti
anti-
infectant and anti-cancer
cancer activity, apart from being the source of various other pharmaceutically useful
compounds [5].

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Many bioactive compounds are isolated from Actinomycetes are source of diverse clinical effects and
have a great important applications in human medicine. The need for novel and safe antibiotics is key
challenge for the pharmaceutical industry now days. The discovery of new antibiotics represents
screening of more and more microbes. These microorganisms may have capability to produce some of the
most important medicines ever developed .The resistance problem demands to discover new
antibacterial agents effective against resistant pathogenic bacteria and fungi. So, we need to screen more
and more actinomycetes from different habitats for antimicrobial activity in the hope of getting some new
actinomycetes strains that produce novel antibiotics, which have not been discovered yet and are active
against drug-resistant pathogens [1].

NATURE AND SOURCES OF ACTINOMYCETES

The actinomycetes have been found in different niches such as soil, air, fresh water, oceans and variety of
materials like manure, compost, plant residues and food products. Actinomycetes are found in aquatic
environments and consequently in drinking water system. Only a couple of examination has been carried
out to understand the diversity of actinobacteria in the extreme environments, ecological role and their
adaptation. In order to find novel bioactive compounds of pharmacological and industrial relevance,
actinobacteria have been isolated from exotic and unexplored locations such as desert marine and
wetland areas [6].
Soil actinomycetes or Terrestrial habitat
Actinomycetes constitute a major component of the microbial population in most soils. It was estimated
that actinomycetes commonly obtained from soil at the ratio of almost 1 million per gram and over
twenty genera have been isolated from soil. It was found that 95% of the isolates belonged to
streptomycetes. Environmental factors influence most the type and population of actinomycetes present
in the soil. Most of the isolate obtained from actinomycetes behave as neutrophiles in culture, with their
growth range in between pH 5.0 to 9.0 and an optimum pH around 7.0. The pH is a major environmental
factor determining the distribution, availability and activity of soil actinomycetes. Neutrophiles are
present less in acidic soils whereas acidophilic and acidoduric streptomycetes are found numerous in
acidic soils. For the most part actinomycetes found in the research facility carry on as mesophiles, with
their ideal development temperature at 25 to 30oC. Numerous mesophilic actinomycetes are dynamic in
compost [7].

AQUATIC HABITAT
Fresh water Actinomycetes
Actinomycetes are abundant in fresh water lakes. They are also found in sewage. Various members of
genera Actinoplanes, Micromonospora, Rhodococcus, Streptomyces and the endospore-forming
Thermoactinomycetes have been isolated from freshwater habitats. Majority of these actinomycetes most
probably are wash-in from land and accumulated in fresh water habitats. The presence of Rhodococcus
coprophilus a coprophilic species in lakes is believed due to wash in of contaminated herbivore dung. The
presence of Streptomyces in freshwater habitat is because of their spores being continuously washed into
rivers and lakes [7].
Marine Actinomycetes
Microbial diversity constitutes an infinite pool of novel chemistry, making up a valuable source for
innovative biotechnology [8]. The recent estimates suggest that the culturability of microorganisms in
marine sediments (0.25). The marine environment is a source of interesting research for new species and
a promising source of pharmaceutically important compounds [9]. Since environmental conditions of the
sea are extremely different from terrestrial conditions it is felt that marine actinomycetes may have
different characteristics from terrestrial actinomycetes and therefore might produce novel bioactive
compounds and new antibiotics [8]. The isolation of rare actinomycetes warrants suitable isolation
procedures including the use of appropriate selective media containing macromolecules like casein, chitin
and humic acid for promoting growth of rare actinomycetes present in the samples and simultaneously
suppressing and hindering contaminant bacterial fungal colonies . Actinomycete genera identified by
cultural and molecular techniques from different marine ecological niches include Actinomadura,
Actinosynnema, Amycolatopsis, Arthrobacter, Blastococcus, Brachybacterium, Corynebacterium, Frankia,
Frigoribacterium, Geodermatophilus, Gordonia, Kitasatospora, Mycobacterium, Nocardioides, Nocardiopsis,
Nonomurea, Micromonospora, Micrococcus, Microbacterium, Salinispora, Solwaraspora, Streptomyces,
Williamsia Streptosporangium, Tsukamurella, Dietzia, Psuedonocardia, Rhodococcus, Saccharopolyspora,
Turicella, Serinicoccus and Verrucosispora [10].

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Actinomycetes from plant

Actinomycetes isolate from various medicinal plants part (leaves, fruits, twig) of Catharanthus rosea,
Calotropis procera, Brassica sps., Eugenia caryophyllus and Emblica officinalis (CR1, CR2, CR3, BS1, BS2,
BS3, BS4 and BS5) are 8 actinomycetes species isolated from plant parts [11]. Novel Substances,
Trehangelins Found from the Metabolites of Plant-Derived Rare Actinomycete Polymorphospora rubra
K07-0510 and after Purification of this compound eventually identified three new compounds, which
were named trehangelin A, B, C [12].
Actinomycetes from extreme environments
Actinomycetes are also present in extreme type of habitats. Alkalophilic actinomycetes (Streptomyces and
Nocardiopsis are the dominant genera) are present in alkaline soils (pH 10-12) surrounding mineral
springs. Isolated Saccharomonospora halophila, a halophilic actinomycete with optimum growth at 10%
NaCl from marsh soil Modestobacter multiseptatus, psychrophilic strains with optimum growth at
temperature 11-13oC was isolated from transantarctic mountain soils. An obligate psychrophilic
actinomycetes, Cryobacterium psychrophilum, with optimum growth temperature 9-12oC and did not
grow at temperature higher than 18oC was isolated from Antartica soil. Other than that, acidophilic
actinomycetes have also been isolated from acidic forest and peat soils, mainly Streptomyces and
Micromonospora. Few rare thermotolerant actinomycetes isolated from desert soils of Mojave Desert,
California belonged to genera Microbispora, Nocardia, Microtetraspora, Amycolaptosis, Actinomadura and
Saccharothrix [7].
Actinomycetes identification and characterization
Isolation and identification proof is mandatory before finding the novel characteristic features of any
microbial isolates. With the advancement in genomics, the complexity of microbial world is largely
understandable. In such manner, recent advancements in microbial systematics have led to a ‘polyphasic
taxonomic approach’ which intends to produce all phenotypic, genotypic and phylogenetic information of
a microbial taxon. The prevalent conventional strategies are not adequate to give a complete draft for
microbial taxonomy as these conventional system describe only shape, colour, size, staining properties,
motility, host-range, pathogenicity and absorption of carbon sources. However, Microorganisms present
in the environment can be enumerated, isolated and characterised by various culture dependent classical
techniques such as pour plate and spread plate methods followed by Gram's staining and biochemical
tests to decipher their physiological characteristics. Based upon colony morphology on growth medium,
microscopic observation and biochemical tests isolated bacteria can be assigned to specific genera [13].

Figure 1 (Courtesy: [14], [5] and [15])

Phenotyping
Phenotypic procedures are generally not meant for discrimination among strains of different species.
Determination of unique characteristics of a microorganism allows study of colonization or cross-
infection and enables the establishment of phylogenetic relationships. Biochemical tests alone usually
allow species identification but may also help distinguish among strains of organisms [14].
Genotyping
The identification of DNA analyses aimed that genome of every individual is unique and is basic to all
organisms that reproduce sexually, differences may occur because the offspring inherits different alleles

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from either parent [14]. For identification and characterization of any important biological organism
(prokaryotic or eukaryotic), nucleic acid based molecular approach is required to be the most powerful
methodology [5]. Actinomycetes and its identification based on bacterial sequences of 16S ribosomal DNA
has been started by isolating DNA of 16S rDNA and implying the polymerase chain reaction (PCR) and
[16, 17] Universal Primers was used to amplify the 16S rRNA gene of actinomycetes. For sequencing
reactions, DNA sequencer is used [5]. The Amplification of polymorphic DNA through specific selection of
annealing sites of primer by DNA fingerprinting and differences in the primer-binding sites and between
existence sites lead to synthesis of amplified DNA fragments (amplimers) which vary in length. Terms
such as amplification fragment length polymorphism, interrepeat PCR, DNA amplification fingerprinting,
arbitrarily primed PCR, Restriction fragment length polymorphism (RFLP) and random amplification of
polymorphic DNA (RAPD) are used [15]. Closely related homologs were recognized through using Basic
Local Alignment Tool (BLAST) program and the evolutionary relationships in the sequences were
depicted by constructing a phylogenetic tree for this dendrogram/ phylogram was constructed [18].
Advanced molecular technologies in identification and characterization of Actinomycetes
Different types of physical and chemical pretreatment strategies have been formulated for isolating
desirable rare actinomycetes genera. The utilization of these genus oriented strategies in modern
screening programs has given a significant revelation of new bioactive compounds. These methods
valuable to circumvent the problem of recharacterization of known bioactive molecules and to help in
screening of novel compounds and use of the advanced molecular strategies should make feasible the
discovery of novel pharmaceutical and modern industrial important product [5].

HIGH THROUGHPUT SCREENING (HTS)

The Screening of multiple samples against highly characterized targets unlike the largely cell-based
systems of the past, where efforts were less tailored on the molecular aspects conversely, concluded that
screening compounds using a cell-based biological approach could save three years and more than $300
million of the cost of developing a novel drug [19]. The most noticeable and promising technique for
enzymatic characterization of any microbial population is High Throughput Screening (HTS) strategy
however this strategy is expanded very little for actinomycetes. The HTS strategy is essentially utilized in
drug discovery related field of science mainly chemistry and in biology and some portion of the important
applications of HTS strategy in microbial innovation are all well adopted HTS based strategy; proteomics
approach is starting at now reporting its well acknowledgment in the method for finding the microbial
worlds. High-throughput screening involves screening large libraries of small metabolites against a
particular target [5].
An effective HTS screening system should start with a vigorous assay for the targets. In a study, has
estimated that 100,000 results per system per day are likely to be dwarfed in the future because of the
evolving capabilities of high-throughput screening. This technology is, simultaneously, cross-fertilized by
advances in both automation and bioinformatics [20].
Fluorescence activated cell sorting (FACS)
The requirement of sensitive, high-throughput technologies for the directed evolution of new enzymes
has spurred the development of a new generation of library screening formats. Fluorescence activated
cell sorting (FACS) is a technology that can rapidly separate the cells from suspension dependent on size
and the color of their fluorescence [21]. It is essentially represented with some fluorescent substrates that
are quite certain for a particular enzyme is utilized in the experiment. The positive fluorescence assigns a
definite biocatalytic activity of the clone and the technology is effectively drawn in for desired clones
arranging from a genomic DNA library [5].
Microfluidic FACS (mFACS)
A successful utilization of microfluidic FACS (mFACS) chips in prokaryotic system was published in a
reputed journal on 1999. Escherichia coli cells expressing green fluorescent protein recognized and
separated from a background of nonfluorescent E. coli cells which encouraged a considerable
enhancement of micron-sized fluorescent bead populations with various colors. This detachment was
also confirmed the viability of the bacteria after extraction from the sorting device. This device can be
adopted useful for Actinomycetes cell isolation from a complex microbial population where the device
will be functioned as remain solitary device of an integrated microanalytical chip. Enzyme-fluorescence
technology is Gel MicroDrop technology which is essentially based on identification of positive clones
specific to particular enzymes by catching the fluorescence produced because of catalytic breakdown of
biotinylated substrate by the clone [5].

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Metagenomic approach
Metagenomic approach is another old yet fascinating methodology principally manages the preparation
of a clonal library from the metagenome acquired from extreme habitats (ocean beds, arid regions,
stratosphere, hot stream areas and other) taking into consideration of inability of developing of actual
microbes under laboratory conditions. Despite the fact that this procedure additionally ensured the quick
screening approach by exploring the bioactive potential for unculturable microbes however it limits the
opportunity of regular advantage mainly exhibiting the very low or no expression of desired gene [22].
Substrate-induced gene expression screening (SIGEX)
Most encouraging and pioneering methoodology can be investigated for Actinomycetes characterization
are metagenomic DNA rearranging, Coupling classical functional screens of metagenomic libraries with
innovative approaches are substrate-induced gene expression screening (SIGEX) and pre-amplification
inverse-PCR (PAIPCR) that present learning of useful metagenomics for a specific microflora [5]. With
SIGEX, qualities perhaps engaged with benzoate and catechol degradation have been found in a
groundwater metagenomic library. Despite the limitations of SIGEX, which can only recognized genes that
are effectively induced in a heterologous host, the strategy is extremely high throughput since vast
number of clones can be screened in moderately short timescales [23].
Green fluorescent protein (GFP)
HTS strategy additionally ensures the advancement of technology with the space of rate, parallel
execution and financial to the screening protocol. It comprises of drop based microfluid stage carries a
complex system governing a small foot print chip with a variety of insoluble substrates specific for the
enzyme of interest. Reporter gene technology is another part of the advancement of screening technique
which deals with simplicity and sensitivity of reporter enzyme i.e., Green fluorescent protein (GFP) have
made easier detection of genes in host systems [5].

WHOLE GENOME SEQUENCING APPROACH IN IDENTIFICATION AND CHARACTERIZATION OF

ACTINOMYCETES
Although molecular and advanced technologies have incredible role to blossom up the knowledge by
exploring the Actinomycetes, yet there are few ambiguities still remain related to genomics controlling
the entire mechanics of microbe’s activity. In such cases whole genome sequencing (WGS) approach out
from single cells has made a scientific achievement, which open the whole molecular and biochemical
potential of uncultured microbe from a complex environment. The nucleotide sequence is determined
with an automated sequencing instrument [5]. Instruments of DNA sequencing depends on the
modification of dideoxynucleotide chain terminator chemistry in which the sequencing primer is labeled
at 5’ end with anyone of the four fluorescent dyes. Each fluorescent dye represents one of the four
nucleotides and thus four different annealing and extension reactions are performed. At last, the four sets
of arrangements are combined, concentrated and loaded in a single well on a polyacrylamide gel. During
electrophoresis the fluorescently labeled products are excited and the relating signal is automatically
detected. In this way, the subsequent information is processed into a final sequence with the aid of
computer software. Despite the fact that the ultimate technique for identification, DNA sequencing is
highly expensive and requires a high degree of specialized competency. In this manner, the future
sequencing technique should be simplified and automated further for their appropriateness. At present
the genomes of approximately 200 microbes have been sequenced completely [15].

SIGNIFICANCE OF ACTINOMYCETES
Antibiotics
Actinomycetes have been known as the greatest producer of antibiotics. Almost two third of the total
antibiotics are obtained from actinomycetes [24]. There are almost 45% of 23,000 bioactive microbial
metabolites produced by actinomycetes and the most frequent producers, the Streptomyces species
produce 7,600 compounds. The products from rare actinomycetes in 1970 were only 5%. In that
gathering Streptoverticillium, Micromonospora, Nocardia, Streptosporangium, Saccharopolyspora,
Actinoplanes, and Actinomadura, species are the most successive makers; every one create a few several
antibiotics. Antifungal agent chitinase is produced by actinomycetes that suppress plant pathogenic fungi
and mosquitoes and they may also used in production of single-cell protein, estimation of fungal biomass,
morphogenesis, medical application and degradation of fish wastes, etc. Natural strategies utilized on
huge scale in finding new anti-microbials from actinomycetes. The importance of actinomycetes in
antibiotic production has stimulated many aspects of basic research on these microorganisms. A range of
useful actinomycete antibiotics were reported [25].

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Table.1 Actinomycetes antibiotics and their biological property

Antibiotic Producer Biological Property
Aburamycin Streptomyces aburaviensis Active on gram positive and gram
negative bacteria
Actinomycetin Streptomyces albus and Active on gram positive bacteria
Streptomyces sp.
Actinomycin Streptomycetaecae Older chemotherapy drug
Actinorubin Streptomyces sp. Active on gram positive, negative
bacteria and mycobacteria
Actinoxanthine Streptomyces globisporus Active on gram positive bacteria
Alkavin Streptomyces sp. Active on variety of bacteriophages
Alboverticillin Streptomyces sp. Active on fungi and yeast
Amphotericin-A Streptomyces sp. Active against fungi
Amphotericin-B Streptomyces nodosus Active against fungi
Antibiotic of Streptomyces sp. Active on gram positive and negative
Chandrasekhar bacteria
Carcinomycin S. carcinomycicus, Streptomyces Active on gram positive bacteria and
sp. and Streptomyces Fungi
gannmycicus
Chloramphenicol Streptomyces venezuelae, Active on gram positive and negative
Streptomyces Bacteria
phaeochromogenes var.
chloromyceticus and
Streptomyces omiyaensis
Cyclohexamide Streptomyces griseus, Active on variety of plant pathogens
Streptomyces sp. and
Streptomyces noursei
Enteromycin Streptomyces albireticuli Active on gram negative bacteria
Erythromycin Saccharopolyspora erythraea, Antibacterial and Active on gram
Streptomyces erythreus positive bacteria,
mycobacteria and Corynebacterium
Framycetin Streptomyces lavendulae Active on gram positive and negative
bacteria
Geomycin Streptomyces xanthophacus Active on gram positive and negative
bacteria
Gentamycin Micromonospora sp. Antibacterial against gram-
negativemicroscopic organisms
Hygromycein Streptomyces hygroscopicus and S. Active on gram positive, negative
noboritoensis bacteria and mycobacteria
Hygromycein- B Streptomyces hygroscopicus Active against gram positive and
gram negative bacteria and
Mycobacteria
Kanamycin-A Streptomyces kanamyceticus Active on gram positive, negative
bacteria and mycobacteria
Kanamycin-B Streptomyces kanamyceticus Active on gram positive, negative
bacteria and mycobacteria
Lavendulin Streptomyces lavendulae Active on gram positive, negative
bacteria and mycobacteria
Litmocidin Nocardia cyanca Active on gram positive, negative
bacteria and mycobacteria
Levomycin Streptomyces sp. Moderate activity against gram
positive and gram negative bacteria
and Mycobacteria
Matamycin Streptomyces matensis Active on gram positive bacteria
Melanosporin Streptomyces melanosporus Active on gram positive bacteria and
Fungi
Miamycin Streptomyces sp. Active on gram positive bacteria

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Miramycin Streptomyces mirabilis Active on gram positive and gram

negative bacteria

Moldin Streptomyces Active on gram positive bacteria

phaeochromogenes
Neomycin Streptomyces fradiae, S. Active on gram positive and gram
albogriscolus and Streptomyces sp. negative bacteria

Nocardicin A Nocardia uniformis Antibacterial

Novomycin Streptomyces Active on gram positive and negative
roscochromogenes bacteria
Nucleocidin Streptomyces calvus Active on gram positive and negative
bacteria and mycobacteria
Nystatin Streptomyces noursei Antifungal against organisms
partcularly fungi especially Candida
sp.
Oligomycins Streptomyces sp. Active on filamentous fungi
Paromomycin Streptomyces rimosus Active on gram positive and negative
bacteria and mycobacteria
Perimycin Streptomyces coelicolor Active on yeasts and filamentous
fungi
Picromycin Streptomyces fellas and Active on gram positive bacteria
Streptomyces sp.
Puromycin Streptomyces alboniger Active on gram positive bacteria
Pyridomycin Streptomyces pyridomyceticus Active against mycobacteria
Rhodomycetin Streptomyces griseus Active on gram positive bacteria
Rifamycin Amycolatopsis mediterranei Antibacterial against M. tuberculosis
Ristocetin Amycolatopsis lurida Antibacterial against Streptococcus
sp.
Rubromycin Streptomyces collinus Active on gram positive bacteria
Ruticin Streptomyces sp. And Active on gram positive and negative
S. rutgerscnsis bacteria
Sistomycosin Streptomyces viridosporus Active on yeasts and filamentous
fungi
Spinosyns Saccharopolyspora spinosa Insecticidal
streptomycin Streptomyces griseus, S. bikiniensis Antibacterial against gram- positive
and S. olivaceus and gram-negative bacteria
Streptothricin Streptomyces lavendulae Active on gram positive and negative
bacteria
Streptozotocin Streptomyces achromogenes Active on gram positive and negative
bacteria
Teicoplanin Actinoplanes teichomyceticus Antibacterial against gram- positive
bacteria
Telomycin Streptomyces sp. Active on gram positive bacteria
Tetracycline Streptomyces aureofaciens Antibacterial activity
Tetrin Streptomyces sp. Active on filamentous fungi and yeast
Vancomycin Amycolatopsis orientalis Antibacterial against Streptococcus
sp., mycobacteria and some
spirochetes
Xanthomycins Streptomyces sp. And Very active on gram positive and
S. rutgerscnsis negative bacteria
Xanthothricin S. albus and Streptomyces sp Moderate activity against gram
positive and gram negative bacteria
and Mycobacteria
Zaomycin Streptomyces zaomyceticus Active against gram positive bacteria
[Source: 26, 27 and 1]

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Actinomycetes in plants
In disease control
For the most part actinomycetes in soil have a place with the family Streptomyces and just about 60% of
organically dynamic aggravates that have been created that are utilized as a part of the farming are
started from them [28]. Mycostop a commercial item, in light of S. griseoviridis K16 and S. lydicus
WYEC108 can control root rots and wilt diseased caused by Pythium spp. Fusarium spp., Rhizoctonia spp.
and Phytophthora spp. [29].
Production of plant growth hormone (indole-3-acetic acid)
Actinomycetes mostly used in the analysis of bioactive compounds. A few animal groups shape secondary
metabolites, hostile to helminthic compounds, against tumor specialists and anti-infection agents [30].
Free-living species of actinomycetes have additionally been concerned in the improvement of plant
growth by production of plant growth-producing substances like auxins and gibberellin-like compounds.
Indole-3-acetic acid (IAA), which regulates many basic cellular processes including cell division,
elongation and differentiation, is the principal form of auxin [31].
Biogenic synthesis of metal nanoparticles from actinomycetes
The biosynthesis of metal nanoparticles occurs intracellularly or extracellularly. According to in
actinomycetes, intracellular reduction of metal ions occurs on the surface of mycelia along with
cytoplasmic membrane leading to the formation of nanoparticles. Different metal nanoparticles such as
silver, gold, zinc, and copper synthesized by using actinomycetes showed antimicrobial activity against a
wide range of microbes including multidrug-resistant bacteria and fungi [32].
The mechanism of action of metal nanoparticles involves three mechanisms. Firstly, metal nanoparticles
bind with cell membrane and disturb its power functions, such as permeability and respiration. Silver
nanoparticles may cause depletion of intracellular ATP by rupture of plasma membrane or by blocking
respiration in association with oxygen and sulfhydryl groups on the cell wall to form RS- S-R bonds
leading to cell death. Secondly, silver nanoparticles are able to penetrate into the bacterial cell membrane,
interact with sulfur-containing and phosphorus-containing compounds, such as DNA, and cause damages
inside it [33]. Thirdly, the silver nanoparticles release silver ions, which may contribute to the bactericidal
activity of metal nanoparticles. It is believed that DNA loses its replication ability, and cellular proteins
become inactivated after interaction with silver nanoparticles. The higher concentration of silver
nanoparticles has shown to interact with cytoplasmic components and nucleic acids [34].
Secondary Metabolites secreted by Actinomycetes
Soil habitats and marine environments samples have been used to isolate novel actinomycetes [25]. The
secondary metabolites produced by actinomycetes serve as the sources of life saving environments and
have a broad spectrum of biological activities; e.g. antibacterial (streptomycin, tetracycline,
chloramphenicol), antifungal (nystatin), antiviral (tunicamycin), antiparasitic (avermectin),
immunosuppressive (rapamycin), antitumor (actinomycin, mitomycinC, anthracyclines), Cancer
(doxorubicins, daunorubicin, mitomycin and bleomycin), enzyme inhibitory (clavulanic acid) and
diabetogenic (bafilomycin, streptozotocin), transplant rejection (cyclosporine and rapamycin) and high
cholesterol (statins such as lovastatin and mevastatin [35].
Actinomycetes not only play a great role in the biological activities in addition to this Actinomycetes
species, are makers of clinically valuable antitumor medications, for example, Anthracyclines
(Aclarubicin, Daunomycin and Doxorubicin), Aurelic acids (Mithramycin), Peptides (Bleomycin and
Actinomycin D), Enediynes (Neocarzinstatin), Antimetabolites (Pentostatin), Carzinophilin, Mitomycins
and others. Actinomycetes are not capable for their powerful remedial exercises or essential organic
exercises yet additionally for the use of pharmacokinetic properties required for clinical advancement
[35].
In the start of the anti-toxin time the contagious (penicillin, griseofulvin) and bacterial(Gramicidin)
species were in intrigue, yet after the revelation of streptomyces species the more consideration swings
to streptomyces species, the antibiotics discovered from streptomyces were streptomycin and later
chloramphenicol, tetracyclines and macrolides. In Fifties and sixties Majority of antibiotics almost 70%
were reported from streptomyces and 45% of the presently known metabolites almost about 10000
compounds were still isolated from the various actinomycetales species, 34% of them were from
streptomyces and 11% of them from rare actinomycetes species produce 7600 compounds (74% of all
actinomycetales), although the typical Actinomycetes represent 26%, altogether 2500 compounds [36].
The representation of rare actinomycetes products in 1970 was only 5%. In the gathering Nocardia,
Streptoverticillium, Micromonospora, Streptosporangium, Actinoplanes, Saccharopolyspora and
Actinomadura, species are the most continuous makers; every deliver a few many antibiotics [37].

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Table.2 Number of actinomycetales species producing bioactive microbial metabolites.

Actinomycetales species No. Actinomycetales species No.
Streptomycetaceae: Thermomonosporaceae:
Streptomyces 8000 Actinomadura 345
Streptoverlicillium 258 Saccharothrix 68
Kitasatosporia 37 Microbispora 54
Chainia 30 Actinosynnema 51
Microellobosporia 11 Nocardiopsis 41
Nocardioides 9 Microtetraspora/Nonomuria 26/21
Micromonosporaceae: (Actinoplanetes) Thermomonospora 19
Micromonospora 740 Micropolyspora/Faenia 13/3
Actinoplanes 248 Thermoactinomyces 14
Dactylosporangium 58 Thermopolyspora 1
Ampullariella 9 Thermoactinopolyspora 1
Glycomyces 2 Mycobacteriaceae: (Actinobacteria)
Catenuloplanes 3 Nocardia 357
Catellatospora 1 Mycobacterium 57
Pseudonocardiaceae: Arthrobacter 25
Saccharopolyspora 131 Brevibacterium 17
Amycalotopsis/Nocardia 120/357 Proactinomyces 14
Kibdellosporangium 34 Rhodococcus 13
Pseudonocardia 27 Other (unclassified) species:
Amycolata 12 Actinosporangium 30
Saccharomonospora 2 Microellobosporia 11
Actinopolyspora 1 Frankia 7
Streptosporangiaceae: (Maduromycetes) Westerdykella 6
Streptosporangium 79 Kitasatoa 5
Streptoalloteichus 48 Synnenomyces
4
Spirillospora 11 Sebekia
3
Planobispora 10 Elaktomyces 3
Kutzneria 4 Excelsospora
3
Planomonospora 2 Waksmania 3
Alkalomyces 1
Catellatospora 1
Erythrosporangium 1
Streptoplanospora 1
Microechinospora 1
Salinospora 1
[Source: 36, 37]

Actinomycetes as source of Agroactive compounds

Kasugamycin is a bactericidal and fungicidal metabolite discovered from Streptomyces kasugaensis [26].
The inhibitor of protein biosynthesis in microorganism’s occure due to this antibiotic, and its toxicological
properties are excellent. Hokko Chemical Industries build up a production process to advertise the
systemically active kasugamycin for control of rice blast Pyricularia oryzae Cavara and bacterial
Pseudomonas diseases in several crops.
The isolated Polyoxin B and D were metabolites of Streptomyces cacaoivar. Asoensis and in 1965 by
classified it as a new class of natural fungicides. They play role in the fungal cell wall synthesis by
specifically inhibiting chitin syntheses. Polyoxin B has application against a number of fungal pathogens
in fruits, vegetables and ornamentals. Polyoxin D is marketed by several industries to control rice sheath
blight caused by Rhizoctonia solani Kùhn [26].
Antifungal metabolite mildiomycin isolated from a culture of Streptoverticillium rimofaciens Niida was
reported in 1978, also by Takeda scientists [38]. Mildiomycin antibiotics is strongly active against several

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 Mathur et al

powdery mildews on various crops, Furthermore compounds mentioned are agroactive compounds
isolated from actinomycetes and microbial screening and chemistry techniques have been until recently
the main tools to discover new agroactive compounds [39].
Actinomycetes as Biopesticide Agents
Microorganisms including bacteria, fungi, nematodes and viruses that are antagonistic to insects are
accounted as strategies to biologically control them. Actinomycetes play a great role in the biological
control of insects through the production of insecticidally active compounds against the house fly Musca
domestica [40]. The mortality rates were measured very high almost reaching up to 90% after
actinomycetes treatments at larval and pupal stages and Chitinase enzyme is extremely necessary within
the biological control of insects and the largest chitinase activity among bacteria has been determined in
species of Streptomyces, Serratia, Vibrio and Bacillus [41].
Enzyme production from Actinomycetes
Physiological, Biochemical and Molecular characteristics in actinomycetes followed by metabolic pathway
yield a variety of biologically active enzymes. The varieties of enzymes secreted by actinomycetes are
chitinase (eg. Streptomyces viridificans), cellulases(eg. Thermonospora spp.), proteases (Nocardia spp.),
peptidases, Xylanases (Microbispora spp.), ligninases (Nocardia autotrophica), amylases
(Thermomonospora curvata), sugar isomerases (Actinoplanes missouriensis), hemicellulase, pectinase and
keratinase [42].

Table.3 Applications of enzymes produced from actinomycetes.

Enzymes Application References
1. Amylase I. In fermentation [43]
II. Food industry
III. In textile and paper industry
2. Catalase I. Used as an Antioxidant [26]
II. In Dairy industry
III. In cold Sterilization of beer
3. Cellulases I. In animal feed industry [44,26]
II. In biomechanical pulping
III. In laundry
4. L- asparaginase I. In Bone marrow treatment [45,46,47 and 48]
II. In stem cell transplant
III. In treatment of acute leukemia
5. Lipase I. In oleochemical [49,50]
II. In detergent industry
III. Pharmaceutical industries
IV. In diagnostic setting
6. Urease I. In wine industry
II. Analysis in blood and urine
III. Analysis of heavy metal content in waste [26]
water and soil
7. Proteases I. In cancer treatment [51]
II. Protect against clots
III. Used as Anti-inflammation
8. Chitinase I In plant resistance against [52,53]
fungal pathogen
II In biochemical industry
III In drug delivery and wound
healing
[Source: 7, 26]

Actinomycetes in bioremediation/biodegradation
The pesticides degradation are responsible because of actinomycetes with various different chemical
structures, including organochlorines, s-triazines, triazinones, carbamates, organophosphates,
organophosphonates, acetanilides, and sulfonylureas [7]. Petroleum hydrocarbons are widely used as
chemical compounds and fuel in our daily life. Greater use results, petroleum now one of the most serious
contaminants of large soil surfaces and finally are considered as a major environmental problem [54].

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 Mathur et al

Several ways in which hydrocarbons degraded in the environment and one such mechanism through
which they can be removed from the environment is bioremediation. Bioremediation is the use microbes
to degrade harmful pollutants to harmless substances. Some reports on Streptomyces flora indicated that
they could play a very important role in degradation of hydrocarbons. Actinomycetes have numerous
such properties that make them utilized for application in bioremediation of soils contaminations. They
play role in the recycling of organic carbon and are being able to degrade complex polymers. In some
reports in a few reports Actinomycetes are having more preference among the degraders Actinomycetes
species are having the capacity to live in oil domain. So we can apply these microorganisms in
Bioremediation to deduct oil contaminations. Numerous strains may be able to solubilise lignin and
debase lignin-related compound by creating cellulose-and hemicellulose-degrading proteins and
extracellular Peroxidase [26].
Actinomycetes in Biocorrosion
Corrosion is a main reason of pipe failure and of high costs in gas pipelines [55]. Biocorrosion is
characterized as a sarcastic harm started by the immediate or aberrant exercises of microorganisms
Antimicrobial substance (AMS) delivered by a Streptomycetes strain having its action against a vigorous
bacterium B. pumilus LF-4, and sulfate-diminishing bacterium D. alaskensis NCIMB 13491 known to be
engaged with biofilm arrangement and biocorrosion [26]. One of Strain 235 was identified that belong to
S. lunalinharesii species was initially isolated from a Brazilian soil. This strain was beforehand known as
maker of bioactive mixes against microscopic organisms and parasites [56]. The antimicrobial activity
was seen at different pH, chemicals and temperature but not seen with Proteinase K and trypsin. The
antimicrobial substance are of proteic nature, has advertised for use in oil making plants, demonstrated
its strength within the sight of a few synthetic chemicals, solvents, and at various temperature and pH
values [57].
Nitrogen-fixing actinobacteria Frankia
Frankia is a species of actinomycetes in the family Frankiaceae that fix nitrogen, both are advantageous
and free-living oxygen consuming, while most rhizobia don't [58]. The filamentous gram-positive Frankia
sp. recommends the significant gatherings of nitrogen-fixing symbionts have obtained mechanism for
nitrogen fixation from various transformative starting points [59]. The very first successful isolation
of Frankia was reported recently from Comptonia peregrina root nodules. Till now, we have 200 strains of
Frankia have been isolated from many, despite the fact that not all, actinorhizal plant species.
Phylogenetic examinations uncovered that Frankiae shape an intelligent clade inside actinobacteria.
The Frankia family of actinobacteria and their host plants is form symbiotic relationships with various
species and fixed 15% of the world's nitrogen [60].
Volatile Organic Compounds (VOCs)
Actinomycetes, especially the genus Streptomyces, are well characterized for their capability of producing
a variety of secondary metabolites such as antibiotics [61]. Despite this, there has been little systematic
investigation of the production of volatile organic compounds by these organisms, geosmin and much
attention has been paid to the production of off-odor, musty, aroma compounds produced by these
organisms, mainly geosmin (trans-1, 10-dimethyl-trans-9-decalol) and 2-methylisoborneol, due to the
detrimental effects of these compounds on the quality of fresh water sources and aquaculture-raised fish
(wood et al., 1983). Streptomycetes are characterized by a complex secondary metabolism. They make
more than 66% of the clinically valuable anti-microbials of normal source (e.g. neomycin,
chloramphenicol) [62]. The odor of freshly turned soil is that the results of geosmin, a volatile organic
compound obtained from actinomycetes. Geosmin is additionally made by some cyanobacteria and
produces an earthy taste in drinking water [26].

CONCLUSION
Indirect 23,000 bioactive secondary metabolites delivered by microorganisms have been accounted for
and more than 10,000 of these compounds are created by actinomycetes. A few pharmaceutical
organizations utilized microbial natural items as one of the significant source of novel medications.
Analysts have been going ahead to find more novel metabolites with potential remedial application
particularly from actinomycetes. Further, just little data is accessible on the actinomycetes. Ongoing
discoveries from culture-dependent and culture free techniques have exhibited that indigenous
actinomycetes exist in the seas and are broadly conveyed in various marine biological systems. These
marine actinomycetes deliver diverse kinds of new secondary metabolites. Diversity and novelty
tremendously among in actinomycetes present in marine environments and several new methods have
been used to detach novel actinomycetes from various conditions and natural surroundings include
Different molecular approaches such as genetic fingerprinting, metagenomics, metaproteomics, 16S r

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 Mathur et al

RNA, genus specific primers, RAPD, RFLP, Proteomics and bioinformatics tools are vital for discovering
and characterizing the vast actinomycetes diversity. For novel medication conveyance, researchers still
adventure the synthetic and biological variety from different actinomycetes for successful disclosure of
novel strain in cost effective manner and these techniques useful to circumvent the problem of
recharacterization of known bioactive molecules and to help in screening of novel compounds. Use of the
above mentioned strategies should make feasible the discovery of novel pharmaceutical and industrial
important product from actinomycetes.

CONFLICT OF INTEREST
The authors have no conflicts of interest

ACKNOWLEDGEMENT
The authors are grateful to acknowledge all the colleagues and Jaipur National University, Jaipur,
Rajasthan, India for carried out work in the School of Life Science, Main Campus by providing necessary
facilities in the laboratory for this study.

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CITATION OF THIS ARTICLE

V Mathur, I Jadhav, R Vasniwal, D Shrivastava and K Jadhav. Ubiquity, Significance and the Molecular Techniques used
for deciphering Actinomycetes. Bull. Env. Pharmacol. Life Sci., Vol 9[2] January 2020 : 164-177

BEPLS Vol 9 [2] January 2020 177 | P a g e ©2020 AELS, INDIA