• Title/Summary/Keyword: liquefying enzyme

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Studies on Cellulase -Part 1. Isolation of Cellulase Forming Microorganisms and the Properties of Crude Enzymes- (Cellulase에 관(關)한 연구(硏究) -(제1보(第一報)) Cellulase 생성균(生成菌)의 분리(分離)와 조효소(粗酵素)의 제성질(諸性質)-)

  • Chung, Dong-Hyo
    • Applied Biological Chemistry
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    • v.11
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    • pp.109-117
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    • 1969
  • Out of some 400 strains of Microorganisms, cellulase forming organisms was isolated from night soil during the course of screening tests. Two strains, Ku-3371 and Ku-4383 were found capable of producing cellulase in the shaking culture. General properties of the crude enzyme were as the following results. 1. The optimum pH values on CMC-saccharifying, CMC-liquefying and filter paper disintegrating activities were 4.0 to 5.5. 2. The stable pH range was within 3.5 to 6.5, 3. The optimum temperature was $40-45^{\circ}C$, the thermal stability was below $50^{\circ}C$ except on paper disintegrating activity and completely inactivated at $70^{\circ}C$. 4. Dialyzed crude enzyme was activated by $Mn^{2+}\;and\;Co^{2+}$ repectively but $Hg^{2+}$ was strong inhibitor.

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Isolation of an Agarolytic Bacteria, Cellvibrio mixtus SC-22 and The Enzymatic Properties (한천분해세균 Cellvibrio mixtus SC-22의 분리 및 효소적 특성)

  • Cha, Jeong-Ah;Kim, Yoo-Jin;Seo, Yung-Bum;Yoon, Min-Ho
    • Journal of Applied Biological Chemistry
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    • v.52 no.4
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    • pp.157-162
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    • 2009
  • An agar-liquefying bacteria (SC-22), which produces a diffusible agarase that caused agar softening around the colony was isolated from Daecheong lake in Korea. Chemotaxanomic and phylogenetic analyses based on 16S rRNA gene sequences revealed the strain was classified as Cellvibrio mixtus SC-22. The isolate SC-22 showed maximal extracellular agarase activity with 58.5 U/mL after 48 h cultivation in the presence of 0.2% agar. It was observed that the isolate produced two kinds of extracellular and three kinds of intracellular isoenzymes. The major agarase was purified from the culture filtrate of agarolytic bacteria by ammonium sulfate precipitation, anion exchange and gel filtration column chromatographic methods. The molecular mass of the purified enzyme was estimated to be 25 kDa by SDS-PAGE. The optimum pH and temperature of the purified enzyme were pH 7.0 and $50^{\circ}C$, respectively. The agarase activity was activated by $Fe^{2+}$, $Na^+$ and $Ca^{2+}$ ions while it was inhibited by $Hg^{2+}$, $Mn^{2+}$ and $Cu^{2+}$ at 1 mM concentration. The predominant hydrolysis product of agarose by the enzyme was galactose and disaccharide on TLC, indicating the cleavage of $\beta$-1,4 linkage in a random manner. The enzyme showed high substrate specificity for only agar and agarose among various polysaccharides.

Characterization of Endochitosanases-Producing Bacillus cereus P16

  • Jo, Yu-Young;Jo, Kyu-Jong;Jin, Yu-Lan;Jung, Woo-Jin;Kuk, Ju-Hee;Kim, Kil-Yong;Kim, Tae-Hwan;Park, Ro-Dong
    • Journal of Microbiology and Biotechnology
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    • v.13 no.6
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    • pp.960-968
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    • 2003
  • A bacterial isolate showing a strong endochitosanase activity was isolated from soil and then characterized. The isolate was identified and designated as Bacillus cereus P16, based on morphological and biochemical properties, assimilation tests, cellular fatty acids pattern, along with 16S rRNA gene sequence. The optimized medium for producing extracellular chitosanase in a batch culture contained 1% tryptone, 0.5% chitosan, and 1% NaCl (pH 7.0). Powder chitosan and tryptone served the best as carbon and nitrogen sources, respectively, for the chitosanase production. Chitosanase activity was the highest when culture was completed at $37^{\circ}C$ among various temperatures ($20-42^{\circ}C$) tested in a shaking incubator (200 rpm). The levels of chitosanase activity in the culture fluid were 2.0 U/ml and 3.8 U/ml, respectively, when incubated in a flask for 60 h and in a jar fermenter for 24 h. The culture supernatant showed a strong liquefying activity on the soluble chitosan. The viscosity of 1% chitosan solution, that was incubated with the culture supernatant, was rapidly decreased, suggesting the secretion of endochitosanolytic enzymes by P16. The culture fluid revealed six endo-type chitosanase isozymes, two major (38 and 45 kD), and four minor (54, 65, 82, and 96 kD) forms by staining profile. The crude enzymes were very stable, and full activity was maintained for 4 weeks at $4^{\circ}C\;or\;-20^{\circ}C$ in the culture supernatant, suggesting a highly desirable stability rate for making an industrial application of the crude enzymes. The supernatant also cleaved the insoluble chitosan powder, but the hydrolysis rate was much lower. The enzymic degradation products of chitosan contained $(GlcN)_n$ (n=2-8). The concentration of chitosan in the reaction mixture of the crude enzyme affected the chitooligosaccharides composition of the hydrolysis products. When the higher concentration of chitosan was used, the higher degree of polymerized chitooligosaccharides were produced. By comparison with other commercial chitosanase preparations, P16 was indeed found to be a valuable enzyme source for industrial production of chitooligosaccharides from chitosan.

Effect of Condiments on the Microflora, Enzyme Activities and Taste Components of Traditional Kochujang during Fermentation (양념류를 첨가한 전통고추장의 숙성중 미생물, 효소활성 및 맛 성분의 변화)

  • Kim, Dong-Han
    • Korean Journal of Food Science and Technology
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    • v.33 no.2
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    • pp.264-270
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    • 2001
  • In order to improve the quality of traditional kochujang, condiments like garlic and onion were added to kochujang and their effect on microbial characteristics, enzyme activities and taste components were investigated during fermentation. Viable cells of yeasts in the kochujang decreased with the increasing ratio of garlic. However, aerobic and anaerobic bacterial counts did not show any remarkable differences in the garlic or onion added kochujang, they decreased slowly after $6{\sim}10$ weeks of fermentation. The activity of liquefying amylase in kochujang decreased slowly during $2{\sim}10$ weeks of fermentation, but that of saccharifying amylase increased remarkably at $14{\sim}18$ weeks. Amylase activities increased at the late period of fermentation in garlic or onion added kochujang. Protease activities of kochujang were strong in the middle of fermentation, but they decreased by addition of garlic or onion. The major free sugars in kochujang were glucose and fructose, and their contents increased as the ratio of garlic increased. The major organic acids in kochujang were succinic, malic and oxalic acid, and they also decreased by addition of garlic and onion. The contents of total free amino acids and amino nitrogen were the highest in 2% garlic added kochujang. The major free amino acid were glutamic acid, aspartic acid, proline, alanine, leucine, isoleucine and arginine. Serine, glutamic acid, lysine and arginine contents in increased by adding garlic or onion.

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Effect of Red Pepper Varieties on the Microflora, Enzyme Activities and Taste Components of Traditional Kochujang during Fermentation (고추품종을 달리한 전통식 고추장의 숙성중 미생물, 효소활성 및 맛성분의 변화)

  • Shin, Dong-Hwa;Kim, Dong-Han;Choi, Ung;Lim, Mi-Sun;An, Eun-Young
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.26 no.6
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    • pp.1050-1057
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    • 1997
  • Microbial counts, enzyme activities and taste components of traditional kochujangs prepared with the powders from 4 different varieties of red pepper, were investigated during 90 days fermentation for the industrial production of traditional kochujang. The viable cell counts of anaerobic bacteria in kochujangs did not change remarkably during fermentation, however, aerobic bacterial counts showed a rapid increase up to 90 days of aging. The yeasts in all kochujang samples increased until 60 days of aging and than decreased. After 90 days of aging, the count of aerobic bacteria in Kumtop kochujang was higher than those of others. The activities of liquefying amylase decreased during the aging, but those of saccharogenic amylase increased at 60 days of aging. The activities of neutral protease were higher than those of acidic protease, and increased during the middle and last period of aging. The major free sugars of kochujang were maltose and glucose, and their contents were higher in Hongkwang kochujang. The major organic acids of kochujang were succinic, formic and citric acid, followed by lactic acid. Succinic acid content in kochujang decreased during fermentation, whereas formic and citirc acids were increased. The major free amino acids were serine, proline, glutamic acid, aspartic acid and alanine. Kumtop kochujang contained the highest amount of total free amino acids. Among the nucleotides and related components in kochujang, cytidine-5-monophosphate was the most abundant component at the begining of aging period, while hypoxanthine increased remarkably during fermentation. Hanwang kochujang was higher in the content of nucleotides than others. Capsaicin contents of kochujang decreased during aging and those of Jangter kochujang was higher than that of others. Sensory evaluation showed that Jangter kochujang was significantly better than Kumtop kochujang in overall acceptability, but there were no appreciable differences in color and flavor.

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Optimization of Demucilage Process of Opuntia ficus-indica var. saboten Fruit using High Hydrostatic Pressure Enzyme Dissolution (고압 효소 액화 장치를 이용한 백년초 점질물 분해 공정의 최적화)

  • Im, Sungbin;Lee, Hyungjae;Shim, Jae-Yong;Kim, Tae-Rahk;Kim, Dae-Ok
    • KSBB Journal
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    • v.30 no.2
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    • pp.63-68
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    • 2015
  • This study aimed to develop and optimize a demucilaging process of Opuntia ficus-indica var. saboten (OFI) fruit to increase its usability as functional food ingredient and food additive. Viscozyme and Novozym 33095 as multienzyme complex having a broad spectrum of carbohydrases and pectolytic enzymes, respectively, were used in enzymatic dissolution along with high hydrostatic pressure liquefaction. To optimize the liquefaction process using high hydrostatic pressure liquefying extractor, response surface methodology with 3-factor central composite design was employed with reaction factors such as temperatures (25, 32, 40, 48, and $55^{\circ}C$), pressures (20, 40, 60, 80, and 100 MPa), and times (15, 30, 45, 60, and 75 min). At optimum conditions ($25^{\circ}C$, 100 MPa, and 58.275 min) for high hydrostatic pressure liquefaction process, the processed OFI fruit juice was predicted to have viscosity at 2.917 poise, partly due to the release of free sugars such as fructose and glucose detected using HPLC-ELSA system. The results above suggests that the OFI fruit juice with decreased viscosity may be used for various manufacturing processes of food, beverage, ice cream, and cosmetics.

Studies on the Brewing of Kochujang (Red Pepper Paste) with the Addition of Mixed Cultures of Yeast Strains (효모혼용(酵母混用)에 의한 고추장의 양조(釀造)에 관(關)한 연구(硏究))

  • Lee, Taik-Soo;Yang, Keel-Ja;Park, Yoon-Joong;Yu, Ju-Hyun
    • 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.

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Characteristics of Jochung by Wet-Milled Rice Flour and Steamed Rice (습식 미분과 증미로 제조한 쌀조청의 특성)

  • Lee, Jung-Eun;Choi, Yoon-Hee;Cho, Mun-Gyeong;Park, Shin-Young;Kim, Eun-Mi
    • The Korean Journal of Food And Nutrition
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    • v.25 no.3
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    • pp.637-643
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    • 2012
  • This study was performed to increase the utilization of rice and improve the productivity of jochung, traditional food in Korea. Two kinds of jochung products were prepared from steamed rice(SR) and wet-milled rice flour(WRF) by rice cultivated from 2006 to 2010. It is common to add a liquefying enzyme for rice liquefaction(4 $m{\ell}$/1,000 g rice, at $90{\sim}95^{\circ}C$, 3 h) and saccharogenic enzyme with malt(45 g/1,000 g rice, at $55{\sim}57^{\circ}C$, 6 h). In order to evaluate the quality characteristics of jochung, producing rate, pH, solidity, reducing sugar, dextrose equivalence(D.E.), viscosity, total phenolic compound, color value and sensory evaluation were carried out. In terms of the producing rate of jochung, WRF jochung was produced about 7.4% much more than SR jochung. There was no difference in producing rate between the jochung cultivated from 2006 to 2010 rice. The pH varied from 4.86~5.66, solidity was 79.48~82.28%. Color L value was 25.82~27.92, a value of 1.28~2.81, b value were 2.98~4.33. The results of sensory evaluation for jochung, as a whole, received higher score than for the commercial product(Daesang Co., Ltd, Seoul, Korea), overall acceptability score was the highest in the 2008SR. Reducing sugar, dextrose equivalence(D.E.) and total phenolic compounds were determined to be higher WRF jochung than SR jochung, while viscosity was lower WRF jochung than SR jochung. These results are thought to be due to increased surface area of rice by milling. SR jochung manufacturd by wet-milled rice flour will increase the producing rate for jochung, thereby saving manufacturing time and costs.

Changes in Microflora and Enzymes Activities of Traditional Kochujang Prepared with Various Raw Materials (담금원료에 따른 전통식 고추장의 숙성 중 미생물과 효소력의 변화)

  • Shin, Dong-Hwa;Kim, Dong-Han;Choi, Ung;Lim, Mi-Sun;An, Eun-Young
    • Korean Journal of Food Science and Technology
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    • v.29 no.5
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    • pp.901-906
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    • 1997
  • In order to reproduce and improve quality of traditional kochujang, various raw materials were added to prepare kochujang by replacing part of the glutinous rice. Chemical composition, microbial characteristics and enzyme activities were investigated during fermentation. Crude protein and salt contents of kochujang did not change significantly during fermentation, but moisture contents increased linearly. The pH and titratable acidity of kochujang changed little in garlic added group. The viable cell counts of aerobic bacteria and yeasts in the kochujang increased until 60 days of fermentation and then decreased slowly except for the garlic added group in which they increased during the last period of fermentation. Aerobic bacterial count did not show any remarkable differences among the samples and slowly decreased after 60 days of fermentation. The activities of liquefying and saccharifying amylases decreased until 45 days, but increased at 60th day. Acidic protease activities of each group were strong during the initial period, but neutral protease showed the highest activity from the 30 to 45 days of fermentation. Protease activities increased by addition of soy sauce, Chinese matrimony vine and purple sweet potato.

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A Study on the Preparation of Traditional Andong sikhe with Lactic Acid Bacteria (유산균을 이용한 전통안동식혜의 제조방법에 관한 연구)

  • Choi, Cheong;Son, Gyu-Mok
    • Journal of the Korean Society of Food Culture
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    • v.7 no.3
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    • pp.259-270
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    • 1992
  • The changes in life style today appear many ways. Many housewives turn away from home preparation of the time consuming traditional foods, such as 'Andong sikhe'. The importance, however, of succeeding the traditional cuisines is getting appreciated widely nowadays. This study aimed to investigate the preparation of Andong sikhe by use of pure culture inoculation and the improvement of storage stability by the addition of stabilizers to the product. Lactobacillus delbreuckii was selected for the pure culture inoculation in the fermentation. The changes in chemical composition such as total acidity, sugar content, amino acid and various forms of nitrogen during fermentation were determined. The changes in pH of the product, the enzyme activities and the population of lactic acid bacteria were also followed in the process of fermentation. The Lactobacillus dominated in the beginning of the fermentation but the Streptococcus out numbered the former as the fermentation proceeded. The crude protein content increased up to the 4th day of fermentation but slowly decreased there after. The pH of the product rapidly decreased to 4.2 by the 2nd day of fermentation. The total acidity reached to the 0.38% by the 2nd day of fermentation and kept on increasing slowly during the fermentation. The free sugar consisted of 6 kinds including maltose and one unknown sugar. The amino form nitrogen increased up to 38.5mg% at the 2nd day of fermentation and the product tasted best at this time. The ammonia form nitrogen, water soluble and salt soluble protein decreased during fermentation. Proline and aspartic acid were the two major free amino acids. The free methionine increased while the free lysine decreased in the process of fermentation. The major amino acids of water soluble and salt soluble protein were glutamic acid and aspartic acid. The arginine content of salt soluble protein increased as the fermentation proceeded. Linoleic, palmitic and oleic acid were the three major fatty acids and occupy 90% or more of the total fatty acids. The activities of acid protease and liquefying amylase reached to the maximum at the 4th day of fermentation while those of saccharogenic amylase and lipase reached to the peak at the 2nd day of fermentation.

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