• 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.

• Biotechnology and Bioprocess Engineering:BBE
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• 제8권5호
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• pp.303-305
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• 2003

Scale-up of hirudin production from Saccharomyces cerevisiae from bench-scale to pilot-scale was carried out based on constant volumetric oxygen transfer coefficient (K$\sub$L/a). Fed-batch mode of cultivation using step-wise feeding strategy of galactose was employed for the production of hirudin in a 30-L and a 300-L pilot-scale fermentor. The final hirudin concentrations were achieved 390 mg/L and 286.1 mg/L, and the volumetric productivities were 80.4% and 90.7% with the 30-L and 300-L fermentors, respectively, compared to the productivity of the 5-L bench-scale fermentor.

• 한국식품영양과학회지
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• 제28권2호
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• pp.348-352
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• 1999

We have intended to accelerate the secretion of human lysozyme(HLY) with permeabilizing agents from the cultivated cells of the recombinant Saccharomyces cerevisiae. The five agents CaCl2, Tween 80, ethanol, Triton X 100, and cetyltrimethylammonium bromide(CTAB) were used as permeabilizing agents. Treatments of the yeast cell with CaCl2, Tween 80, and ethanol were effective to increase the secretion from the yeast cells. Especially, treatment of 10% ethanol increased the extracellular HLY activity by 38.6% at 30oC for 48 h in culture broth. But Triton X 100 and CTAB unexpectedly didn't play a role in increase of HLY secretion. Recovery of a foreign protein by permeabilizing agents is easier than by osmotic shock, and is less expensive than enzymatic digestion.

• KSBB Journal
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• 제17권1호
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• pp.38-43
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• 2002

중금속 결합 단백질인 metallothionein (MT)를 발현하는 CUP1 유전자를 다중으로 함유하는 재조합 Saccharomyces cerevisiae의 균체성장과 중금속 제거특성을 조사하였다. CUP1 유전자를 다중으로 함유하는 재조합 효모는 중금속을 포함한 배지에서의 균체성장과 중금속 흡착능이 중금속에 대하여 내성을 가지고 있는 야생효모나 숙주세포에 비하여 우수하였다. 서로 다른 플라스미드를 함유하는 중금속을 함유하는 배지에서의 균체성장과 중금속 흡착능의 차이는 중금속에 대한 내성과 중금속 흡착에 MT 단백질의 발현 수준이 중요한 영향을 미치는 것으로 추정되었다. 구리를 함유하지 않고 고농도의 아연과 망간을 함유하는 배지에서 재조합 효모는 높은 균체 농도와 중금속 제거능을 나타내었는데, 이는 MT 단백질을 발현하는 CUP1 promoter가 아연과 망간에 의해서도 발현이 유도된다는 것을 알 수 있었다. MT 단백질을 효율적으로 발현하여 재조합 효모에 의한 중금속 흡착을 최대화 할 수 있는 최적의 구리농도는 0.31 mM로 결정되었으며, 비이온계 계면활성제인 Triton X-100은 재조합 효모의 균체성장과 중금속 흡착을 증가시켰지만, 대사 저해제는 균체성장과 중금속 흡착을 모두 저해하였다.

• 생명과학회지
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• 제11권2호
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• pp.190-195
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• 2001

The cuclodextrin glucanotransferase (CGTase) gene of Bacillus macerans was subcloned at the downstream of yeast ADH1 promoter, and then the resulting plasmid pVT-CGTM(9.15 kb) was introduced into the yeast host strain, Saccharomyces cerevisias 2805. The transformed yeast, S. cerevisiae 2805/pVT-CGTM, showed the starch-hydrolyzing activity on the starch-azure plate. The optimal conditions for the CGTase expression were found to be 2% dextrose, initial pH5.5, 3$0^{\circ}C$, and 48hr cultivation. Under this condition, the extracellular CGTase activity reached at 0.53 U/mL, whereas the intracellular activity was about 0.03U/mL. This result indicates that the signal peptide of Bacillus CGTase functioned well in S. cerevisiae.

• KSBB Journal
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• 제13권4호
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• pp.456-460
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• 1998

Fed-batch fermentations were carried out in order to improve the efficiency of hirudin production by recombinant Saccharomyces cerevisiae. A fed-batch fermentation done with the optimized semi-synthetic medium resulted in a maximum hirudin concentration of 342mg/$\ell$ by keeping a galactose concentrations between 10 and 30g/$\ell$ which corresponded to a 11.4-fold increase in hirudin concentration compared with the simple bach fermentation done with the same medium. Comparison of the chromatographic pattern of proteins in the growth medium clearly showed that the use of the semi-synthetic medium is more advantageous for separation of hirudin than the case o fusing the complex medium. Continuous cell recycle fermentation done at dilution rate of 0.1h-1 and an inlet galactose concentration of 100g/$\ell$ yielded a maximum hirudin productivity of 19.1mg hirudin/$\ell$$.$h.

• Biotechnology and Bioprocess Engineering:BBE
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• 제3권2호
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• pp.55-60
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• 1998

Various Saccharomyces cerevisiae strains were transformed with a 2 ${\mu}$-based multicopy expression plasmid, pYIGP, carrying Kluyveromyces marxianus inulinase gene under the control of GAPDH promoter. Among then two strains, SEY2102 and 2805, showed high levels of cell growth and inulinase expression, and were selected to study their fermentation properties on inulin. Jerusalem artichoke inulin was more effective for cell growth (10∼11 g-dry wt./L at 48 hr) and inulinase expression (1.0 units/mL with SEY2102/pYIGP and 2.5 units/mL with 2805/pYIGP) than other inulin sources such as dahlia and chicory. It was also found that maximal ethanol production of 9 g/L was obtained from Jerusalem artichoke inulin at the early stationary phase (around 30 hr), indicating that recombinant S. cerevisiae cells secreting exoinulinase could be used for the simultaneous saccharification of inulin and ethanol fermentation.

• 한국미생물·생명공학회지
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• 제18권3호
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• pp.305-308
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• 1990

Shuttle plasmid vector인 pHN114를 이용하여 Kluyveromyces fragilis의 3-isopropylmalate dehydrogenase 유전자를 cloning 하였다. 그 결과 Saccharomyces cerevisiae의 leu2변이와 E.coli의 leuB변이를 상보하는 두가지 clone 체 pJK104와 pJK106을 얻었다. Restriction mapping 결과 이들은 서로 반대방향으로 삽입되어 있었으며 expression activity는 pJK104가 높았다. pJK104에 삽입된 유전자를 BglII와 SalI으로 끊은 1.6kb fragment를 probe 로 하여 Southern Hybridization 한 결과 유전자의 유래가 Kluyveromyces fragilis 임을 확인하였다.

• 한국미생물·생명공학회지
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• 제23권6호
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• pp.652-658
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• 1995

We attempted simultaneous expression of genes coding for endoglucanase and $\beta $-glucosidase from Pseudomonas sp. by using a synthetic two-cistron svstem in Escherichia coli and Saccharomyces cerevisiae. Two-cistron system, 5'--tac promoter-endoglucanase gene--$\beta $-glucosidase gene-- 3', 5'-tac promoter--$\beta $-glucosidase gene--endoglucanase gene--3' and 5'-tac promoter--endoglucanase gene--SD sequence--$\beta $-glucosidase gene--3, were constructed, and expressed in E. coli and S. cerevisiae. The E. coli and S. cerevisiae contained two-cistron system produced simultaneously endoglucanase and $\beta $-glucosidase. The recombinant genes contained the bacterial signal peptide sequence produced low level of endoglucanase and $\beta $-glucosidase in S. cerevisiae transformants: Approximately above 44% of two enzymes was localized in the intracellular fraction. The production of endoglucanase and $\beta $-glucosidase in veast was not repressed in the presence of glucose or cellobiose. The veast strain contained recombinant DNA with two genes hydrolyzed carboxvmethyl cellulose, and these endoglucanase and $\beta $-glucosidase degraded CMC synergistically to glucose, cellobiose and oligosaccharide. This result suggests the possibility of the direct bioconversion of cellulose to ethanol by the recombinant yeast.

• Journal of Microbiology and Biotechnology
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• 제30권2호
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• pp.296-305
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• 2020

Tricholoma matsutake is an ectomycorrhizal fungus, related with the host of Pinus densiflora. Most of studies on T. matsutake have focused on mycelial growth, genes and genomics, phylogenetics, symbiosis, and immune activity of this strain. T. matsutake is known for its unique fragrance in Eastern Asia. The most major component of its scent is (R)-(-)-1-octen-3-ol and is biosynthesized from the substrate linoleic acid by the sequential reaction of lipoxygenase and peroxide lyase. Here, we report for the first time the biosynthesis of (R)-(-)-1-octen-3-ol of T. matsutake using the yeast Saccharomyces cerevisiae as a host. In this study, cDNA genes correlated with these reactions were cloned from T. matsutake, and expression studies of theses genes were carried out in the yeast Saccharomyces cerevisiae. The product of these genes expression study was carried out with Western blotting. The biosynthesis of (R)-(-)-1-octen-3-ol of T. matsutake in recombinant Saccharomyces cerevisiae was subsequently identified with GC-MS chromatography analysis. The biosynthesis of (R)-(-)-1-octen-3-ol with S. cerevisiae represents a significant step forward.