Synthetic Biology of Thermophiles: Taking Bioengineering to the Extremes?

Vavitsas, Konstantinos; Glekas, Panayiotis D.; Hatzinikolaou, Dimitris G.

doi:10.3390/applmicrobiol2010011

Cited by 9 publications

(4 citation statements)

References 53 publications

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“…The range of optimum activity temperatures for invertase varies widely in the literature. However, thermostable enzymes are generally preferred for industrial applications 32 . Therefore, the invertase reported here is a potential industrial enzyme, considering its optimum temperature and thermal stability.…”

Section: Discussionmentioning

confidence: 99%

Production and characterization of intracellular invertase from Saccharomyces cerevisiae (OL629078.1), using cassava-soybean as a cost-effective substrate

Osiebe,

Adewale,

Omafuvbe

2023

Sci Rep

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The growing global market for industrial enzymes has led to a constant search for efficient, cost-effective methods for their production. This study reports the production of invertase using inexpensive and readily available agro-materials. Starch-digesting enzymes extracted from malted unkilned sorghum were used to hydrolyze cassava starch supplemented with 2% whole soybean. The production of intracellular invertase by Saccharomyces cerevisiae OL629078.1 in cassava-soybean and yeast sucrose broth was compared. The purification and characterization of invertase produced using the low-cost medium were also reported. The results showed that there was a 4.1-fold increase in the units of invertase produced in cassava-soybean medium (318.605 U/mg) compared to yeast sucrose broth medium (77.6 U/mg). The invertase produced was purified by chromatographic methods up to 5.53-fold with a recovery of 62.6%. Estimation of the molecular weight with gel filtration indicated a molecular weight of 118 kDa. The enzyme demonstrated its maximum activity at 50 °C and there was no decrease in its activity following a 1-h incubation at this temperature. At a pH of 5.0, the enzyme demonstrated optimal activity and it maintained over 60% of its activity in the acid range (pH 3–6). The Michalis-Menten constants Km and Vmax of intracellular invertase were 5.85 ± 1.715 mM and 6.472 ± 2.099 U/mg, respectively. These results suggest that Saccharomyces cerevisiae grown on cassava-soybean is a viable, cost-effective alternative for commercial invertase production, which can be explored for biotechnological processes.

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Section: Discussionmentioning

confidence: 99%

Production and characterization of intracellular invertase from Saccharomyces cerevisiae (OL629078.1), using cassava-soybean as a cost-effective substrate

Osiebe,

Adewale,

Omafuvbe

2023

Sci Rep

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“…The survival ability of this thermophilic microorganism is related to its cell structure which has several advantages (Vavitsas et al, 2022);(Sohlenkamp & Geiger, 2016); (Wang et al, 2021) Cell membrane structure (Cebrián et al, 2017); (Parrilli et al, 2022); Chaperonins (Novik et al, 2019); (Villain et al, 2022); (Singh et al, 2021) Structure of DNA gyrase…”

Section: Sourcesmentioning

confidence: 99%

Exploration of Thermophilic Bacteria: Systematic Literature Review

Indriati,

Megahati S.

2023

jppipa, pendidikan ipa, fisika, biologi, kimia

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The selection of enzymes derived from microbes has several advantages compared to those derived from animals and plants. One of the advantages is that the growth of enzymes will occur more quickly in bacteria because the cells rapidly undergo division and cell production will be easier to increase if needed in large quantities and the time needed for enzyme production will be shorter. Thermophilic microorganisms are known as bacteria that are tolerant of high temperatures, but on the other hand these bacteria are also able to survive at these temperatures. Where the purpose of research is to explain exploration of thermophilic bacteria. A review is conducted on the state-of-the-art methods using the preferred reporting items for reviews and meta-analyses (PRISMA) guideline. Bacteria have important economic value in human life. Knowledge in this branch of science is useful in medicine, hygiene, food science and nutrition, agriculture, and industry. Based on the results of macroscopic, microscopic and biochemical characterization, it was found that many bacteria with various types recording to their habitat or place of development, exploraton of thermophilic bacteria was carried out so that they could be useful for the industrial world.

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“…High temperature exerts a fundamental stress on biological systems by destabilizing proteins, membranes, and other molecules (16). Tolerating this stress has driven evolution since life began (17), is relevant to understanding the response of pathogens to fever (18), could be helpful to engineer more efficient cell factories (19), and will become a more common selection pressure as global warming continues (20). One prior study demonstrated the presence of epistasis among mutations acquired under this stress (21).…”

Section: Introductionmentioning

confidence: 99%

Laboratory evolution reveals transcriptional mechanisms underlying thermal adaptation ofEscherichia coli

Rychel,

Chen,

Catoiu

et al. 2024

Preprint

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Adaptive laboratory evolution (ALE) is able to generate microbial strains which exhibit extreme phenotypes, revealing fundamental biological adaptation mechanisms. Here, we use ALE to evolveEscherichia colistrains that grow at temperatures as high as 45.3°C, a temperature lethal to wild type cells. The strains adopted a hypermutator phenotype and employed multiple systems-level adaptations that made global analysis of the DNA mutations difficult. Given the challenge at the genomic level, we were motivated to uncover high temperature tolerance adaptation mechanisms at the transcriptomic level. We employed independently modulated gene set (iModulon) analysis to reveal five transcriptional mechanisms underlying growth at high temperatures. These mechanisms were connected to acquired mutations, changes in transcriptome composition, sensory inputs, phenotypes, and protein structures. They are: (i) downregulation of general stress responses while upregulating the specific heat stress responses; (ii) upregulation of flagellar basal bodies without upregulating motility, and upregulation fimbriae; (iii) shift toward anaerobic metabolism, (iv) shift in regulation of iron uptake away from siderophore production, and (v) upregulation ofyjfIJKL, a novel heat tolerance operon which we characterized using AlphaFold. iModulons associated with these five mechanisms explain nearly half of all variance in the gene expression in the adapted strains. These thermotolerance strategies reveal that optimal coordination of known stress responses and metabolism can be achieved with a small number of regulatory mutations, and may suggest a new role for large protein export systems. ALE with transcriptomic characterization is a productive approach for elucidating and interpreting adaptation to otherwise lethal stresses.

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Synthetic Biology of Thermophiles: Taking Bioengineering to the Extremes?

Cited by 9 publications

References 53 publications

Production and characterization of intracellular invertase from Saccharomyces cerevisiae (OL629078.1), using cassava-soybean as a cost-effective substrate

Production and characterization of intracellular invertase from Saccharomyces cerevisiae (OL629078.1), using cassava-soybean as a cost-effective substrate

Exploration of Thermophilic Bacteria: Systematic Literature Review

Laboratory evolution reveals transcriptional mechanisms underlying thermal adaptation ofEscherichia coli

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