Microbial Biotechnology

Microbial Biotechnology: Basics and Recent Reviews
The terms biodiversity and biological diversity refer to the variety, richness, and complexity of
living things. The biodiversity of microbes came into existence mainly by evolution and natural selection.
Even though there is a wide array of species which are yet to be characterized, we are gripping to
manage and preserve this biodiversity.
Microbes constitute up to billions of species of bacteria, yet only 1-5% of them are known to
human beings and many are still believed to be found. They are incredibly minute and serve to be the
maximum proportion of species exhibiting diversity among living beings in the world. They flourish in
unfavorable pH and temperature ranges, hot springs, ice, hypersaline environments, hydrothermal vent
sites, and other harsh environments. Microbial activities make earth livable and have limitless
commercial applications principally in the field of life science (Arrigo, 2005).
Microbes have been used in beer, wine, acetic acid, and cheese and yoghurt production and
involved in many industries, viz. baking, leather, paper pulp and textile industries (Acharya and
Chaudhary, 2012). A summary of the role of microbial biotechnology for sustainable development of the
ecosystem has been provided in Figure 1.
Energy Improved Food

Animal Breeding Fermented Food
Management Technology
Recombinant Genomics
Pharmaceuticals Proteomics Study
DNA Technology Development
Renewable Biofuels Sustainable

Biorefining
Chemicals Development Environment
Figure 1. Application of microbial biotechnology to maintain the sustainable development of the

ecosystem.
Scope of Microbial biotechnology
Knowing the context of bacteria and its vital role in the field of microbial biotechnology, we
need to understand further the various abilities of the bacteria which can be exploited, majorly the
genetic material utilized for opening a wide array of functions marking the crux of the creation of a
similar yet a newer organism of our interest. A throughput data of bacterial genetics, physiology and
metabolism of millions of gene encoded protein acts as a base for recreating biotechnologically sound
products ultimately leading to the betterment of the society.
Satelliting throughout most of the important microbial aspects of biotechnology, including the
methods put into it and future perspectives, there is no iota of doubt that this era has a potential for
extensive growth. Ranging from genetic engineering to gene therapy, mineral leaching to
bioremediation of toxic wastes and from production of many hormones to microbial fertilizers and
medicines, advances in industrial uses is occurring rapidly.
Genetic Engineering of Microorganisms
Genetic engineering allows scientists to modify the genetic makeup of microorganisms to
enhance desirable traits or confer new functionalities. Techniques such as gene cloning, CRISPR-Cas9,
and synthetic biology have revolutionized microbial biotechnology by enabling the production of
valuable proteins, biofuels, and pharmaceuticals. This section will explore the tools and techniques used
in microbial genetic engineering.
Genetically engineered bacteria (GEBs) are nowadays used as the main body in the recent
advances in most of the bacteriotherapy including the introduction of newer engineering techniques,
administration strategies, performance indicators and biological safety. By the end of the 20 th century,
numerous GEBs are being applied in various aspects of chemical synthesis, food industry, treatment of
diseases and environmental protection. Evidently, Escherichia coli, Lactobacillus and Salmonella species
are the most commonly manipulative bacteria among the GEBs and are used as the structural tools for a
variety of other GEBs.
To date, GEBs have made great achievements in the management of various diseases, such as
infectious diseases, antibiotic-related diarrhea, allergies, and metabolic syndromes, in health care for
daily life (Ma et al., 2017; Mazhar et al., 2020). A broad variety of vaccines in genetically engineered
bacterial format activate majority of the effective immune responses when administered in vivo, such as
Salmonella, Vibrio cholera, Listeria and Neisseria meningitides. Some bacteria like Salmonella,
Clostridium novyi-NT and E. coli are gradually being introduced into the solid tumor cancer therapy.
Therefore, progressing themselves into efficient drug delivery and drug protection.
The basis of microbial genetic engineering involves the specific use of “gene cutting” tools and
editing them further to obtain proper functional genes which exhibits their functions in a repetitive
quality and quantity. Hence, the recombinant genes are administered into newer bacteria with similar
species but with somewhat unique phenotypes.
Enumerating the two major steps of GEB construction; namely, functional gene acquisition (also
known as upstream stage) and heterogeneous expression (downstream stage). The upstream part
involves collection of target genes which are part of a large gene cluster. Thanks to the multiple genome
projects and databases, we can avail and modify the expected bacterial gene. For smaller fragments
(<10kb), DNA can be synthesized directly or by long PCR method (Fahnøe and Bukh, 2019). For segments
more than 50kb, recombination methods, such as CRISPR-Cas method can be applied to alter the gene
using a guide RNA for any kind of deletion or insertion in plasmids or genomes (Li and Elledge, 2012; Luo
et al., 2016; Alberti and Corre, 2019; Strain-Damerell et al., 2021)
Cloning of a gene attains a simple meaning of transferring a particular gene in a vector and
creating a huge number of its copies for a particular purpose. Even though the technique sounds simple,
it includes using complex techniques of transfection, transduction, conjugative transfer, lysogenic
conversion and protoplast fusion. Furthermore, homologous recombination technologies, primarily
homologous recombination, site-specific recombination, transposable recombination, and the CRISPR-
Cas9 technology, allow for the direct integration of target genes into the host chromosome in a
predictable strain.
Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR) and CRISPR- associated
(Cas) proteins constitute an RNA-guided adaptive immune system found in several bacteria and most
archaea. The most impeccable characteristic of CRISPR-Cas-based gene editing is the ability to introduce
heritable genome modifications by programing Cas proteins using guide RNAs, instead of modifying the
editing protein which overcomes the limitation of using ZFNs and TALENs.
The complex process has been distinguished into three stages; Adaptation, crRNA processing
and Interference. Adaptation involves excising of a small fragment of intruder DNA into the CRISPR array
and formation of a new spacer. Cas proteins assist the spacer to be inserted catalytically moving it to the
crRNA processing stage. In this, a long pre-crRNA is transcribed and along with Cas proteins into mature
crRNAs. The last stage shows the cleavage of the intruder DNA or RNA. This cleavage is sequence and
target-specific due to the presence of proteins.
The fitness and economic benefits of an active CRISPR system, as well as the preservation of
degenerated CRISPR-Cas systems in various bacteria, provide promising future study directions. Keeping
in mind about its ability to insert over 50kb of genome in the required target, it enables us to advance
rapidly in most of the aspects of microbial biotechnology including therapeutic, experimental and
diagnostic techniques. Although CRISPR-Cas is the stepping stone of advancement in microbial
biotechnology, there are significant off-targets and a lot of damages (Haapaniemi et al., 2018; Wang et
al., 2020) being analyzed that needs to be overturned in the upcoming future for a better precision and
perfect utilization of this powerful technology.
Figure 2. Generally used recombination technologies from phage infection to CRISPR-Cas9 system
(counter clockwise order) (Liu et al., 2022)
REFERENCES
Acharya S, Chaudhary A (2012) Bioprospecting thermophiles for cellulase production. Braz J Microbiol
43:844–856
Alberti, F., and Corre, C. (2019). Editing streptomycete genomes in the CRISPR/ Cas9 age. Nat. Prod. Rep.
36, 1237–1248. doi: 10.1039/c8np00081f
Arrigo K (2005) Marine microorganisms and global nutrient cycles. Nature 437:349–355 Baldrian P,
Kolařík M, Stursová M, Kopecký J, Valášková V, Větrovský T, Zifčáková L, Snajdr J, Rídl J, Vlček C,
Voříšková J (2012) Active and total microbial communities in forest soil are largely different and
highly stratified during decomposition. ISME J 6(2):248–258
Fahnøe, U., and Bukh, J. (2019). Full-length open reading frame amplification of hepatitis C Virus.
Methods Mol. Biol,1911, 85–91. doi: 10.1007/978-1-4939-8976-8_5.
Haapaniemi, E., Botla, S., Persson, J., Schmierer, B., and Taipale, J. (2018). CRISPR–Cas9 genome editing
induces a p53-mediated DNA damage response. Nat. Med. 24, 927–930. doi: 10.1038/s41591-
018-0049-z
Liu Y, Feng J, Pan H, Zhang X and Zhang Y (2022) Genetically engineered bacterium: Principles, practices,
and prospects. Front. Microbiol. 13:997587. doi: 10.3389/fmicb.2022.997587
Luo, Y., Enghiad, B., and Zhao, H. (2016). New tools for reconstruction and heterologous expression of
natural product biosynthetic gene clusters. Nat. Prod. Rep. 33, 174–182. doi:
10.1039/c5np00085h
M., Elledge, S. J. (2012). SLIC: a method for sequence- and ligationindependent cloning. In Methods Mol
Biol, 852, 51–9. doi: 10.1007/978-1-61779-564-0_5
Ma, B., Pan, Q., and Peppelenbosch, M. P. (2017). Genetically engineered bacteria for treating human
disease. Trends Pharmacol. Sci. 38, 763–764. doi: 10.1016/j. tips.2017.07.001
Mazhar, S. F., Afzal, M., Almatroudi, A., Munir, S., Ashfaq, U. A., Rasool, M., et al. (2020). The prospects
for the therapeutic implications of genetically engineered probiotics. J. Food Qual. 2020, 1–11.
doi: 10.1155/2020/9676452
Newsom S, Parameshwaran HP, Martin L and Rajan R (2021) The CRISPR-Cas Mechanism for Adaptive
Immunity and Alternate Bacterial Functions Fuels Diverse Biotechnologies. Front. Cell. Infect.
Microbiol. 10:619763. doi: 10.3389/fcimb.2020.619763
Strain-Damerell, C., Mahajan, P., Fernandez-Cid, A., Gileadi, O., and Burgess-Brown, N. A. (2021).
Screening and production of recombinant human proteins: ligation-independent cloning,
Structural Genomics, 2199, 23–43. doi: 10.1007/978-1-0716-0892-0_3
Wang, Y., Wang, M., Zheng, T., Hou, Y., Zhang, P., Tang, T., et al. (2020). Specificity profiling of CRISPR
system reveals greatly enhanced off-target gene editing. Sci. Rep. 10, 2269. doi: 10.1038/s41598-
020-58627-x

Microbial Biotechnology

Uploaded by

Copyright:

Available Formats

Microbial Biotechnology

Uploaded by

Document Information

Copyright

Available Formats

Share this document

Share or Embed Document

Sharing Options

Did you find this document useful?

Is this content inappropriate?

Copyright:

Available Formats

Microbial Biotechnology

Uploaded by

Copyright:

Available Formats

Microbial Biotechnology: Basics and Recent Reviews

Energy Improved Food

Renewable Biofuels Sustainable

Figure 1. Application of microbial biotechnology to maintain the sustainable development of the

You might also like