• KSBB Journal
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• 제27권2호
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• pp.127-130
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• 2012

In this study, the effects of pH, temperature, IPTG concentration and substrate (MSG) concentration on gamma-aminobutyric acid (GABA) production in engineered Escherichia coli were investigated. Glutamate decarboxylase and glutamate/GABA antiporter were overexpressed in GABA aminotransferase knock-out strain for GABA production. The result of optimization study showed the GABA bioconversion was optimized at pH 3.5, $30^{\circ}C$, 0.5 mM IPTG, 10 g/L MSG. At this condition, 5.23 g/L of final GABA concentration of was achieved from 10 g/L of MSG, which corresponded to a GABA yield of 85.77%.

• Journal of Microbiology and Biotechnology
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• 제26권8호
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• pp.1348-1357
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• 2016

An enzymatic reaction system was developed and optimized for bioconversion of resveratrol from glucose. Liquid enzyme extracts were prepared from Alternaria sp. MG1, an endophytic fungus from grape, and used directly or after immobilization with sodium alginate. When the enzyme solution was used, efficient production of resveratrol was found within 120 min in a manner that was pH-, reaction time-, enzyme amount-, substrate type-, and substrate concentration-dependent. After the optimization experiments using the response surface methodology, the highest value of resveratrol production (224.40 μg/l) was found under the conditions of pH 6.84, 0.35 g/l glucose, 0.02 mg/l coenzyme A, and 0.02 mg/l ATP. Immobilized enzyme extracts could keep high production of resveratrol during recycling use for two to five times. The developed system indicated a potential approach to resveratrol biosynthesis independent of plants and fungal cell growth, and provided a possible way to produce resveratrol within 2 h, the shortest period needed for biosynthesis of resveratrol so far.

• 한국미생물·생명공학회지
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• 제17권3호
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• pp.219-223
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• 1989

Hydrocortisone의 $\Delta$$^1$-Dehydrogenation에 의한 prednisolone의 생산에서 소포제인 silicone oil과 neolin 302의 영향이 조사되었다. 미생물 전환방법은 pseudo-crystallofermentation 기법에 의해 수행되었다. 스테로이드 입자들은 소포제와의 hydrophobic-hydrophobic interaction에 의해 서로 응집되었다. 이러한 응집 현상에 의해 고체 기질의 용해속도와 비례하는 물질전달 면적이 감소됨으로써, 결국 생물전환속도의 감소를 유발하게 된다. Neolin은 silicone oil에 비해 전환속도에 더욱 좋지 않은 영향을 끼쳤으며, 소포제 농도가 증가할수록 전환속도는 감소하였다.

• Journal of Microbiology and Biotechnology
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• 제28권3호
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• pp.439-447
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• 2018

The aromatic compound p-hydroxybenzoate (PHBA) is an important material with multiple applications, including as a building block of liquid crystal polymers in chemical industries. The cytochrome P450 (CYP) enzymes are beneficial monooxygenases for the synthesis of chemicals, and CYP53A15 from fungus Cochliobolus lunatus is capable of executing the hydroxylation from benzoate to PHBA. Here, we constructed a system for the bioconversion of benzoate to PHBA in Escherichia coli cells coexpressing CYP53A15 and human NADPH-P450 oxidoreductase (CPR) genes as a redox partner. For suitable coexpression of CYP53A15 and CPR, we originally constructed five plasmids in which we replaced the N-terminal transmembrane region of CYP53A15 with a portion of the N-terminus of various mammalian P450s. PHBA productivity was the greatest when CYP53A15 expression was induced at $20^{\circ}C$ in $2{\times}YT$ medium in host E. coli strain ${\Delta}gcvR$ transformed with an N-terminal transmembrane region of rabbit CYP2C3. By optimizing each reaction condition (reaction temperature, substrate concentration, reaction time, and E. coli cell concentration), we achieved 90% whole-cell conversion of benzoate. Our data demonstrate that the described novel E. coli bioconversion system is a more efficient tool for PHBA production from benzoate than the previously described yeast system.

• 한국식품과학회지
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• 제49권1호
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• pp.28-34
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• 2017

본 연구를 통하여 P. aeruginosa PR3를 이용하여 DOD를 생산하기 위해 저가의 기질로서 필리핀 너트유가 효과적으로 사용될 수 있음을 확인하였으며 배지에 첨가되는 여러 영양인자들의 영향을 조사하여 DOD 생산성을 크게 향상시킬 가능성이 있음도 확인하였다. 따라서 DOD 생산에 이용되는 올레산을 식물성오일로부터 별도의 생산과정을 거쳐 생산하지 않고 식물성오일자체를 직접 기질로 사용함으로서 PR3 균주를 이용하여 고부가가치의 DOD를 효율적으로 생산할 수 있다는 것을 확인하였다.

• Journal of Microbiology and Biotechnology
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• 제27권1호
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• pp.77-83
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• 2017

Lignocellulosic biomass represents a potentially large resource to supply the world's fuel and chemical feedstocks. Enzymatic bioconversion of this substrate offers a reliable strategy for accessing this material under mild reaction conditions. Owing to the complex nature of lignocellulose, many different enzymatic activities are required to function in concert to perform efficient transformation. In nature, large multienzyme complexes are known to effectively hydrolyze lignocellulose into constituent monomeric sugars. We created artificial complexes of enzymes, called rosettazymes, in order to hydrolyze glucuronoxylan, a common lignocellulose component, into its cognate sugar ${\small{D}}$-xylose and then further convert the ${\small{D}}$-xylose into ${\small{D}}$-xylonic acid, a Department of Energy top-30 platform chemical. Four different types of enzymes (endoxylanase, ${\alpha}$-glucuronidase, ${\beta}$-xylosidase, and xylose dehydrogenase) were incorporated into the artificial complexes. We demonstrated that tethering our enzymes in a complex resulted in significantly more activity (up to 71%) than the same amount of enzymes free in solution. We also determined that varying the enzyme composition affected the level of complex-related activity enhancement as well as overall yield.

• Journal of Microbiology and Biotechnology
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• 제33권1호
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• pp.1-14
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• 2023

Polyethylene terephthalate (PET) is a plastic material commonly applied to beverage packaging used in everyday life. Owing to PET's versatility and ease of use, its consumption has continuously increased, resulting in considerable waste generation. Several physical and chemical recycling processes have been developed to address this problem. Recently, biological upcycling is being actively studied and has come to be regarded as a powerful technology for overcoming the economic issues associated with conventional recycling methods. For upcycling, PET should be degraded into small molecules, such as terephthalic acid and ethylene glycol, which are utilized as substrates for bioconversion, through various degradation processes, including gasification, pyrolysis, and chemical/biological depolymerization. Furthermore, biological upcycling methods have been applied to biosynthesize value-added chemicals, such as adipic acid, muconic acid, catechol, vanillin, and glycolic acid. In this review, we introduce and discuss various degradation methods that yield substrates for bioconversion and biological upcycling processes to produce value-added biochemicals. These technologies encourage a circular economy, which reduces the amount of waste released into the environment.

• Journal of Microbiology and Biotechnology
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• 제12권3호
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• pp.518-521
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• 2002

Poly(3-hydroxyalkanoate) copolyesters containing both carbon-carbon double and carbon-carbon triple bonds were produced by Pseudomonas oleovorans grown in mixtures of 10-undecynoic acid (10-UND($\equiv$)) and 10-undecenoic acid (10-UND(=)). The PHA content in the dry cells was usually 40 wt%. The bioconversion yield of ($10-UND({\equiv})$) to PHA by P. oleovorans was remarkably enhanced from 1% to over 24% as the fraction of 10-UND(=) in the carbon substrate mixtures increased from 0 to 50%. These values were higher than those obtained when P. oleovorans was grown in the same molar mixtures of ($10-UND({\equiv})$) and nonanoic acid (NA), indicating that 10-UND(=) was more efficient than NA as a cosubstrate in inducing cometabolic PHA production.

• 한국미생물·생명공학회지
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• 제16권3호
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• pp.187-192
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• 1988

Cholesterol로부터 미생물전환 방법에 의한 4-androstene-3,17-dione(AD)의 생성에 대해 연구를 수행했다. 발효액중에 cholesterol의 용해도를 증가시키기 위해 ethanol을 용매로 사용하였는데, ethanol 농도가 2%(v/v)까지는 세균성장이 크게 저해되지 않았다. 미생물전환은 pH를 7.0으로 조절하고, 초기대수 증식기에 cholesterol을 첨가했을 때 효율적으로 AD가 생성되었다. AD 생성을 높이기 위해 cholesterol 첨가방법을 여러 가지로 변환시켰다. 즉, 최종 cholesterol 농도를 0.1% 하여 ethanol에 녹여 간헐적으로 첨가했을 때 가장 높은 수율을 얻었다. Cholesterol을 세번(전체 3g/$\ell$) 첨가했을 때 최종 전환수율이 65%에 도달한 반면, 같은 양의 cholesterol (3g/$\ell$)을 한번에 넣었을 때 40%의 생성수율을 얻었다.

• 한국생물공학회:학술대회논문집
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• 한국생물공학회 2001년도 추계학술발표대회
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• pp.529-532
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• 2001

Korean food waste, one of the abundantly available but environmentally problematic organic wastes in Korea, was utilized as solid-substrate by fungal strain Aspergillus niger ATcC 6275 for the production of enzymemixture containing amylase, cellulase and xylanase. The enzyme mixture can be used as high value-added animal feed. Solid-state fermentation method yielded a 84-fold enhancement in xylanase activity compared with submerged fermentation method. The effect of incubation period, incubation temperature, pH of medium, moisture content, inoculum size and enrichment of the medium with nitrogen and carbon sources were observed for optimal production of these enzymes The optimal amylase activity of 33.10 U/g, cellulase activity of 24.41 U/g, xylanase activity of 328.84 U/g were obtained at 8 days incubation with 50%(w/w) soy bean flake, with incubation temperature of $25^{\circ}C$, pH of 6.38, optimal moisture content of 55% and with inoculum size of $3.8{\times}10^6$spore/g. Enzyme activities were enhanced when ImM $CaSO_4$, 2% Malt extract and 2% galactose were added as mineral, nitrogen and carbon enrichment respectively.