• Journal of Microbiology and Biotechnology
• /
• v.28 no.6
• /
• pp.909-916
• /
• 2018

Previously, a cytosolic trehalase (TreH) from the hyperthermophilic archaeon Sulfolobus acidocaldarius was reported; however, the gene responsible for the trehalase activity was not identified. Two genes, saci_1816 and saci_1250, that encode the glycoside hydrolase family 15 type glucoamylase-like proteins in S. acidocaldarius were targeted and expressed in Escherichia coli, and their abilities to hydrolyze trehalose were examined. Recombinant Saci_1816 hydrolyzed trehalose exclusively without any help from a cofactor. The mass spectrometric analysis of partially purified native TreH also confirmed that Saci_1816 was involved in proteins exhibiting trehalase activity. Optimal trehalose hydrolysis activity of the recombinant Saci_1816 was observed at pH 4.0 and $60^{\circ}C$. The pH dependence of the recombinant enzyme was similar to that of the native enzyme, but its optimal temperature was $20-25^{\circ}C$ lower, and its thermostability was also slightly reduced. From the biochemical and structural results, Saci_1816 was identified as a trehalase responsible for trehalose degradation in S. acidocaldarius. Identification of the treH gene confirms that the degradation of trehalose in Sulfolobus species occurs via the TreH pathway.

• Korean Journal of Microbiology
• /
• v.28 no.2
• /
• pp.120-127
• /
• 1990

Intraspecific fusion frequency between Filobasidium capsuligenum CBS6122-2 ade and met trp or met strains was $2.9\times 10^{-3}$ and $8.5\times 10^{-3}$, respectively. And intergeneric fusion frequency between Folobadidium capsuligenum CBS 4318 (cys his) and Saccharomyces cerevisiae 262-12-1 (hom3 thr1) or X2180-1A (ade thr) was $8.8\times 10^{-6}$ and $9.5\times 10^{-4}$, respectively. Nuclear fusion appears to occru in fusants between intraspecies and intergenera as strongly suggested by DNA content, nuclear staining, comparison of survival rate to UV light and mitotic segregation analysis. It was also found that $\alpha$-amylase and glucoamylase activity from intraspecific hybrids was 1.6-2.1 fold increased when compared with that from thier parents.

• Korean Journal of Food Science and Technology
• /
• v.22 no.3
• /
• pp.296-299
• /
• 1990

As an improved preparation method of Shikhae, a tea-bag system containing malt and amylolytic enzymes was developed in which extraction of malt enzymes and saccharification occured efficiently. The amylolytic activity of the malt was increased by adding the mixture of ${\alpha}-amylase$, glucoamylase and glucoisomerase. Malt and the mixture of enzymes were placed in tea-bag $(16{\times}20cm)$, extracted in water at $30-40^{\circ}C$ for 1-2 hours and followed by saccharification of the cooked rice at $60-70^{\circ}C$ for 3-4 hours. In the conventional Shikhae, content of maltose was about 50% and that of oligosaccharides larger than trisaccharides was about 40% of total sugar. The content of monosaccharides such as glucose and fructose was about 95% and this improved method would be effective for increasing the sweetness and the monosaccharide contents in the product.

• Korean Journal of Agricultural Science
• /
• v.23 no.1
• /
• pp.80-89
• /
• 1996

The expression and secretion of human lactoferrin (hLf) in Sacclnromyces diastaticus were performed. 1. For the secretion of hLf in yeast, recombinant plasmid pYEGLf was constructed using promoter, secretion signal sequence of glucoamylase I gene (STA1) and transcriptional terminator of GAL7 gene. 2. Each correct recombinant plasmid was selected by mini-preparation of plasmid DNA from E coli transformant and restriction enzyme digestion analysis. The selected plasmids, pYEGLf, were transformed into S. diastaticus YIY345 as a expression host, respectively. 3. Western blot analysis using rabbit anti-hLf was carried out to identify expressed hLf. Positive signals were shown in culture supernatant of pYEGLf transformant. 4. About $100{\mu}g-1mg$ of concentrated culture supernatant of positive clone were loaded on paper disc and tested for the antimicrobial activity against E coli. However, no activity was observed. We concluded that this fact results from low concentration of hLf secreted from yeast, compared with the fact that MIC of hLf is as high as $3mg/m{\ell}$. Therefore, the purification of secreted hLf may be require to investigate the antimicrobial activity. From this study, the feasibility of low-cost production of sufficient quantities of human lactofferin for nutritional and therapeutical applications were suggested.

• Korean Journal of Food Science and Technology
• /
• v.25 no.5
• /
• pp.502-509
• /
• 1993

Changes in microflora and enzyme activities of 3 kinds of traditional kochujang were investigated during 6 months of fermentation. Tested kochujang included Sunchang kochujang prepared with glutinous rice, Boeun kochujang prepared with barley and Sachun kochujang prepared with wheat. The pH in Sunchang and Sachun kochujang showed a slighlt decrese during fermentation. In contrast, pH of Boeun kochujang decreased rapidly up to 90 days of fermentation and then leveled off thereafter. The final pH values of Sunchang, Boeun and Sachun kochujang were 4.7, 4.0, and 4.6, respectively. The viable cell counts of aerobic bacteria in Sunchang and Sachun kochujang did not show remarkable changes during fermentation, however, those in Boeun kochujang showed a rapid increase up to 60 days of fermentation and stabilized. On the other hand, the viable cell counts of anaerobic bacteria decreased after 120 days of fermentation. Yeasts were found in different traditional kochujang at different time during the first 60 days of fermentation. It was found that ${\alpha}-$, ${\beta}-$, and glucoamylase activities of Sachun kochujang were higher than those of Sunchang and Boeun kochujang during fermentation. Acidic and neutral proteases showed the highest activity during $30{\sim}60\;days$ and $60{\sim}90\;days$ of fermentation, respectively. Sunchang kochujang showed the highest activity of acidic protease followed in decreasing order by Sachun and Boeun kochujang. On the other hand, Boeun kochujang showed the highest activity of acidic protease followed in decreasing order by Sachun and Sunchang kochujang.

• Microbiology and Biotechnology Letters
• /
• v.24 no.6
• /
• pp.726-732
• /
• 1996

To improve the fermentation characteristics(such as starch-degradability, ethanol tolerance, sugar and high-temperature tolerance) of recombinant haploid yeast Saccharomyces diastaticus K114, hybridization technique was used. The hybridization partner was S. diastaticus 1177 which had good glucoamylase activity and fermentabi- lity. The best hybrid HH64 showed improved ethanol tolerance, sugar and high-temperature tolerance. Especia- lly, the starch-fermentability was significantly improved, since the hybrid produced 1.60% (w/v) ethanol from 4% (w/v) starch, while the recombinant haploid K114 produced 1.30% (w/v) ethanol. The optimum temperature and pH for the starch-fermentation by the hybrid HH64 was 30$\circ$C and 5, respectively. The hybrid yeast HH64 produced 7.5% (w/v) ethanol directly from 20% (w/v) starch.

• The Korean Journal of Food And Nutrition
• /
• v.13 no.2
• /
• pp.134-138
• /
• 2000

An inhibitory activity against ${\alpha}$-glucosidase was identified in extracts of medicinal lichens, Parmelia austrosinensis and P. praesorediosa. The extracts retained almost all of their original activities when treated with heat, acid and alkaline conditions, and some hydrolytic enzymes. Partially purified inhibitor showed strong inhibition against disaccharide hydrolytic enzymes of mammalian and mold origin, but weak or no inhibition against polysaccharide hydrolytic enzymes except glucoamylase. The inhibitors from the two Parmelia sp. showed almost same retention time in HPLC. The inhibitor suppressed elevation of blood glucose level in rats after oral administration of soluble starch or sucrose.

• Archives of Pharmacal Research
• /
• v.8 no.2
• /
• pp.67-75
• /
• 1985

To find emylase inhibitors produced by microorganisms from soil, a strain which had a strong inhibitory activity against bacteria .alpha.-amylase was isolated from the soil smaple collected in Seoul. The morphological and physiological characteristics of this strain on several media and its utilization of carbon sources showed that it was one of Streptomyces specties according to the international Streptomyces Project method. The amylase inhibitor of this strain was purified by means of acetone precipitation, adsorption on Amberlite XAD-2, and column chromatography on Amberlite CG-50 and SP-Sephadex C-25. The inhibitor was stable at the pH range of 1-10 and at 100.deg.C for half an hour, and had inhibitory activities against other amylases such as salivary .alpha.-amylase, pancreatic .alpha.-amylase, fungal .alpha.-amylase and glucoamylase. The kinetic studies of the inhibitor showed that its inhibitory effect on starch hydrolysis by .alpha.-amylase was non-competitive.

• The Microorganisms and Industry
• /
• v.19 no.2
• /
• pp.34-41
• /
• 1993

Industrial strain improvement is concerned with developing or modifying microorganisms used in production of commercially important fermentation products. The aim is to reduce the production cost by improving productivity of a strain and manipulating specific characteristics such as the ability to utilize cheaper raw materials or resist bacteriophages. The traditional empirical approach to strain improvement is mutation combined with selection and breeding techniques. It is still used by us to improve the productivity of organisms in amino acids, organic acids and enzymes production. The breeding of high L-lysine-producing strain Au112 is one of the outstanding examples of this approach. It is a homoserine auxotroph with AEC, TA double metabolic analogue resistant markers. The yield reaches 100 g/l. Besides, the citric acid-producing organism Aspergillus niger, Co827, its productivity reaches the advanced level in the world, is also the result of a series mutations especially with $^60Co{\gamma}$-radiation. The thermostable .alpha.-amylase producing strain A 4041 is the third example. By combining physical and chemical mutations, the strain A 4041 becomes an asporogenous, catabolite derepressed mutant with rifamycin resistant and methionine, arginine auxotroph markers. The .alpha.-amylase activity reaches 200 units/ml. The fourth successful example of mutation in strain improvement is the glucoamylase-producing strain Aspergillus niger SP56, its enzyme activity is 20,000 units/ml, 4 times of that of the parental strain UV-11. Recently, recombinant DNA approach provides a worthwhile alternative strategy to industrial strain improvement. This technique had been used by us to increase the thermostable .alpha.-amylase production and on some genetic researches.

• Korean Journal of Food Science and Technology
• /
• v.27 no.1
• /
• pp.56-62
• /
• 1995

This study was designed to investigate the changes in microflora and enzyme activities of traditional kochujang meju during fermentation for 60 days. The pH of meju decreased continously up to 40 days of aging and then increased slightly thereafter, while the change in titratable acidity showed the opposite trend to that of pH. The viable cell count of aerobic bacteria increared gradually for up to 40 days of fermentation and then decreased slightly thereafter, while that of molds and yeasts showed a rapid increase up to 40 days of fermentation and then leveled off. ${\alpha}-amylase$ activity increased slightly for up to 40 days of meju fermentation and then stabilized. On the other hand, ${\beta}-amylase$ and glucoamylase activities did not show a significant change for up to 20 days of fermentation and then increased rapidly at 40th day of fermentation. Acidic, neutral and alkaline protease activities increased sharply up to 40 days of aging and then decreased significently at 60th day of fermentation. These results suggest that meju fermented for 40 days had the highest quality in terms of the number of microflora and enzyme activity.