• Applied Biological Chemistry
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• v.39 no.3
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• pp.195-199
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• 1996

We have cloned several E. coli sfs genes which stimulate mal gene expression with $crp^{{\ast}1}$). One the genes (pPVC2) was sequenced and potential CRP binding site is located in the upstream of the putative promoter in the regulatory region. In order to investigate the regulation of the sfs1 gene by the cAMP-CRP complex, we have constructed the sfs-lacZ fusion gene in this research. The overall transcriptional stimulations of sfs1 gene in the presence cAMP were confirmed by ${\beta}-galactosidase$ activity and Western blot analysis of sfs1-lacZ fusion gene. Transcriptional regulation by cAMP-CRP was also confirmed by Northern blot analysis. End-labelled DNA of the DNA fragment in sfs1 regulation region were used for gel retardation assay to examine the CRP-DNA complex in the presence of cAMP. Results here indicate that CRP binding site in the regulatory region of sfs1 gene is positive regulator for the expression of sfs1 gene.

• Korean Journal of Food Science and Technology
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• v.4 no.2
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• pp.106-111
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• 1972

The possibility of substituting raw materials for soy sauce by corn gluten was studied by measuring the amylase and proteolytic activities of koji. Also optimum conditions of koji making were determined. It was found that substitution of up to 30% of the defatted bean content (or 15% of the total bean and wheat content) with corn gluten yielded a good quality of soy sauce. Use of more than 15% corn gluten (based on total bean and wheat content) yielded a soy sauce of poor taste and low nitrogen content even though corn gluten has a high nitrogen content. This drop in nitrogen was attributed to the low enzyme activity in koji containing more than 15% corn gluten and the difference in availability of nitrogen in bean compared to corn gluten.

• Korean Journal of Food Science and Technology
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• v.12 no.4
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• pp.313-323
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• 1980

The objective of this experiment was to improve the quality and shortening the aging time of Kochujang by adding mixed starter cultures of yeast strains. Kochujangs were mashed during the summer season with mixed starter cultures of Saccharomyces rouxii, Torulopsis versatilis and Torulopsis etchellsii. Enzyme activities and chemical composition of the Kochujang were determined during the period of aging and their organoleptic values were tested. The maximum activities of liquefying amylase and saccharogenic amylase in the Kochujang were obtained during 20 to 60 days and 20 to 30 days after mashing respectively. The acidic protease activity was reached maximum during 20 to 40 days. All enzyme activities were decreased markedly during the final stage of aging period. Among mixed starter cultures tested, mixed culture of T. versatilis and T. etchellsii shows the highest liquefying and saccharogenic amylase activities. Ethyl alcohol contents in 10 days after mashing were highest in the Kochujang with S. rouxii and T. versatilis, followed in order of S. rouxii and T. etchellsii mixture, T. versatilis and T. etchellsii mixture and control without addition of yeast. But the contents in all sample became similar after 20 days with the level of 2.3 to 2.8% and then decreased gradually. The level of reducing sugar contents was markedly increased during the first 10 days, especially in the batches of T. versatilis and T. etchellsii mixture and control. However, the concentration became similar in all samples after 40 days. The contents of amino nitrogen were increased markedly during the first 10 days then slowly up to 90 days. The rate was high in the Kochujang with T. versatilis and T. etchellsii when compared with others. The organoleptic values of all Kochujang made with addition of yeast starter cultures were superior to control, especially in flavor, taste and color. The Kochujang with T. versatilis and T. etchellsii marked the highest value. The data obtained from this experiment suggests that the quality of Kochujang could be improved by using starter culture of suitable yeast strains according to product characteristics and aging time.

• Korean Journal of Food Science and Technology
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• v.7 no.4
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• pp.229-237
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• 1975

An attempt was made to utilize the enzyme produced by Asp. oryzae as meat tenderizer. The production, purification, and various properties of proteinase produced by Asp. oryzae were investigated. Results obtained are as follow; 1. A strain which had the highest proteolytic activity was selected among 9 Aspergillus species. 2. Culture medium consisted of wheat bran 10g, 2% glucose, 0.03% urea and 0.1% $MgSO_4$ (pH 6.5). Mold was incubated at $30^{\circ}C$ for 3 days. 3. Enzyme extract from culture medium were fractionated with ammonium sulfate and purified by Sephadex G-75 column chromatography. 4. When pH of reaction mixture was controlled, maximal activity of proteinase by Asp. oryzae was obtained at pH 3, pH 6.6, $8.4{\sim}8.5$ and pH 10.0 to 10.5. Those results were interpreted to show that enzyme consists of acid proteinase, neutral proteinase and alkaline proteinase. Enzyme was stable at pH 6 to 10. 5. Opt. temperature for proteinase activity was $50^{\circ}C$, but enzyme was stable up to $40^{\circ}C$. 6. The proteinase was inhibited by $Ag^+$. It was also inhibited by EDTA. 7. When myofibrillar proteins were treated by proteinase from Asp. oryzae, ATPase activities of myofibrillar proteins changed remarkably. Accordingly, it was concluded that proteinase produced by Asp. oryzae were able to be used as meat tenderizer.

• Journal of Life Science
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• v.29 no.4
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• pp.492-498
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• 2019

Previously, three kinds of lipases, lipAD1, lipAD2, and lipAD3, and lipase chaperone, lipBD, of Acinetobacter schindleri DYL129 isolated from soil sample were reported. In this report, three lipase and lipase chaperone were cloned into the pET32a(+) or pGEX-6P-1 vectors for protein expression in Escherichia coli, and named as pETLAD1, pETLAD2, pETLAD3 and pETLBD or pGEXLAD1, pGEXLAD 2, pGEXLAD3 and pGEXLBD, respectively. Protein expression rate was higher in pET system than in pGEX system. Although LipAD1 and LipAD2 were produced as inclusion bodies, their expression levels were high. So LipAD1 and LipAD2 could be solubilized in 8 M urea buffer and purified. LipAD3 and LipBD were overexpressed in soluble form and purified. Those proteins were purified by His-tag affinity chromatography connected in AKTA prime system. The activities of the purified lipases were demonstrated with 1% tributyrin agar plate. After purification, molecular mass was determined with sodium dodecyl sulfate-polyacrylamide gel electrophoresis. LipAD1 showed high activity toward ${\rho}$-nitrophenyl acetate and ${\rho}$-nitrophenyl butyrate, LipAD2 showed high activity toward ${\rho}$-nitrophenyl acetate and ${\rho}$-nitrophenyl myristate, and LipAD3 showed high activity toward ${\rho}$-nitrophenyl acetate, ${\rho}$-nitrophenyl butyrate, and ${\rho}$-nitrophenyl miristate, respectively. Three lipases, LipAD1, LipAD2, and LipAD3, showed optimal reaction at $50^{\circ}C$ using ${\rho}$-nitrophenyl butyrate, as substrate.