• Journal of Microbiology and Biotechnology
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• 제25권4호
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• pp.452-458
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• 2015

The biocatalytic efficiency of recombinant Corynebacterium glutamicum ATCC 13032 expressing the secondary alcohol dehydrogenase of Micrococcus luteus NCTC2665 was studied. Recombinant C. glutamicum converts ricinoleic acid to a product, identified by gas chromatography/mass spectrometry as 12-ketooleic acid (12-oxo-cis-9-octadecenoic acid). The effects of pH, reaction temperature, and non-ionic detergent on recombinant C. glutamiucm whole cell bioconversion were examined. The determined optimal conditions for production of 12-ketooleic acid are pH 8.0, 35℃, and 0.05 g/l Tween80. Under these conditions, recombinant C. glutamicum produces 3.3 mM 12-ketooleic acid, with a 72% (mol/mol) maximum conversion yield, and 1.1 g/l/h volumetric productivity in 2 h; and 3.9 mM 12-ketooleic acid, with a 74% (mol/mol) maximum conversion yield, and 0.69 g/l/h maximum volumetric productivity in 4 h of fermentation. This study constitutes the first report of significant production of 12-ketooleic acid using a recombinant Corynebacterium glutamicum-based biocatalyst.

• Molecules and Cells
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• 제28권4호
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• pp.369-373
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• 2009

Heterologous secretory expression of endoglucanase E (Clostridium thermocellum) and ${\beta}$-glucosidase 1 (Saccharomycopsis fibuligera) was achieved in Saccharomyces cerevisiae fermentation cultures as an ${\alpha}$-mating factor signal peptide fusion, based on the native enzyme coding sequence. Ethanol production depends on simultaneous saccharification of cellulose to glucose and fermentation of glucose to ethanol by a recombinant yeast strain as a microbial biocatalyst. Recombinant yeast strain expressing endoglucanase and ${\beta}$-glucosidase was able to produce ethanol from ${\beta}$-glucan, CMC and acid swollen cellulose. This indicates that the resultant yeast strain of this study acts efficiently as a whole cell biocatalyst.

• 한국생물공학회:학술대회논문집
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• 한국생물공학회 2003년도 생물공학의 동향(XII)
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• pp.101-101
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• 2003

• 한국미생물생명공학회:학술대회논문집
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• 한국미생물생명공학회 2002년도 학술발표대회
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• pp.104-104
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• 2002

• 공업화학
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• 제22권5호
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• pp.486-489
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• 2011

본 연구에서는 대장균 세포 표면에서 발현되는 유기인분해효소(organophosphorus hydrolase; OPH)를 이용하여 난분해성 및 환경독성물질로 알려진 diazinon의 효과적 처리가 이루어졌다. 이 실험에서는 $25^{\circ}C$ 배양 온도와 배지에 0.2 mM ethylenediamine tetraacetate (EDTA) 첨가 조건이 유기인분해 효소 생산에 효과적임을 알 수 있었다. 이 조건에서 성장한 대장균을 이용하여 초음파 파쇄공정이 수행되었을 때, 25, 50 mg/L diazinon는 각각 4.5, $7.2mg/g{\cdot}min$의 diazinon 제거 속도를 나타내었고, 두 농도(25, 50 ppm) 모두 90% 이상 제거 효율을 구할 수 있었다. 따라서 이러한 실험 결과들은 diazinon과 같은 독성 화합물을 친환경적으로 처리할 수 있는 생물학적 처리 시스템으로 활용될 수 있을 것이다.

• 한국생명과학회:학술대회논문집
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• 한국생명과학회 2007년도 제48회 학술심포지움 및 추계국제학술대회
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• pp.63.1-63.1
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• 2007

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• Journal of Microbiology and Biotechnology
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• 제12권3호
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• pp.411-416
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• 2002

Bacillus macerans cyclodextrin glucanotransferase (CGTase) was expressed on the cell surface of Saccharomyces cerevisiae by fusing with Aga2p linked to the membrane-anchored protein, Aga1p. The surface display of CGTase was confirmed by immunofluorescence microscopy and its enzymatic ability to form ${\alpha}$-cyclodextrin from starch. The maximum surface-display of CGTase was obtained by growing recombinant S. cerevisiae at $20^{\circ}C$ and pH 6.0. S. cerevisiae cells displaying CGTase on their surface consumed glucose and maltose, inhibitory byproducts of the CGTase reaction, to enhance the purity of produced cyclodextrins. Accordingly, the experimental results described herein suggest a possibility of using the recombinant S.cerevisiae anchored with bacterial CGTase on the cell surface as a whole-cell biocatalyst for the production of cyclodextrin.

• Journal of Microbiology and Biotechnology
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• 제33권11호
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• pp.1506-1512
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• 2023

Quantitative analysis of adenosine triphosphate (ATP) has been widely used as a diagnostic tool in the food and medical industries. Particularly, the pathogenesis of a few diseases including inflammatory bowel disease (IBD) is closely related to high ATP concentrations. A bioluminescent D-luciferin/luciferase system, which includes a luciferase (FLuc) from the firefly Photinus pyralis as a key component, is the most commonly used method for the detection and quantification of ATP. Here, instead of isolating FLuc produced in recombinant Escherichia coli, we aimed to develop a whole-cell biocatalyst system that does not require extraction and purification of FLuc. To this end, the gene coding for FLuc was introduced into the genome of probiotic Saccharomyces boulardii using the CRISPR/Cas9-based genome editing system. The linear relationship (r² = 0.9561) between ATP levels and bioluminescence generated from the engineered S. boulardii expressing FLuc was observed in vitro. To explore the feasibility of using the engineered S. boulardii expressing FLuc as a whole-cell biosensor to detect inflammation biomarker (i.e., ATP) in the gut, a colitis mouse model was established using dextran sodium sulfate as a colitogenic compound. Our findings demonstrated that the whole-cell biosensor can detect elevated ATP levels during gut inflammation in mice. Therefore, the simple and powerful method developed herein could be applied for non-invasive IBD diagnosis.

• Journal of Microbiology and Biotechnology
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• 제27권1호
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• pp.92-100
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• 2017

Acetoin (AC) is a volatile platform compound with various potential industrial applications. AC contains two stereoisomeric forms: (3S)-AC and (3R)-AC. Optically pure AC is an important potential intermediate and widely used as a precursor to synthesize novel optically active materials. In this study, chiral (3R)-AC production from meso-2,3-butanediol (meso-2,3-BD) was obtained using recombinant Escherichia coli cells co-expressing meso-2,3-butanediol dehydrogenase (meso-2,3-BDH), NADH oxidase (NOX), and hemoglobin protein (VHB) from Serratia sp. T241, Lactobacillus brevis, and Vitreoscilla, respectively. The new biocatalyst of E. coli/pET-mbdh-nox-vgb was developed and the bioconversion conditions were optimized. Under the optimal conditions, 86.74 g/l of (3R)-AC with the productivity of 3.61 g/l/h and the stereoisomeric purity of 97.89% was achieved from 93.73 g/l meso-2,3-BD using the whole-cell biocatalyst. The yield and productivity were new records for (3R)-AC production. The results exhibit the industrial potential for (3R)-AC production via whole-cell biocatalysis.

• Journal of Microbiology and Biotechnology
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• 제16권7호
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• pp.1032-1040
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• 2006

Some Pseudomonas species can grow on styrene as a sole carbon and energy source. From the new isolate Pseudomonas putida SN1, the genes for styrene catabolism were cloned and sequenced. They were composed of four structural genes for styrene monooxygenase (styA and styB), styrene oxide isomerase (styC), and phenylacetaldehyde dehydrogenase (styD), along with two genes for the regulatory system (styS and styR). All the genes showed high DNA sequence (91% to 99%) and amino acid sequence (94% to 100%) similarities with the corresponding genes of the previously reported styrene-degrading Pseudomonas strains. A recombinant Escherichia coli to contain the styrene monooxygenase from the SN1 was constructed under the control of the T7 promoter for the production of enantiopure (S)-styrene oxide, which is an important chiral building block in organic synthesis. The recombinant E. coli could convert styrene into an enantiopure (S)-styrene oxide (ee >99%) when induced by IPTG The maximum activity was observed as 140 U/g cell, when induced with 1 mM IPTG at $15^{\circ}C$.