• Title/Summary/Keyword: Acid protease

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Ruminal Protein Degradation Characteristics of Cell Mass from Lysine Production

  • Seo, S.;Kim, H.J.;Lee, S.Y.;Ha, Jong K.
    • Asian-Australasian Journal of Animal Sciences
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    • v.21 no.3
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    • pp.364-370
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    • 2008
  • Chemical analysis and in vitro studies were conducted to investigate the nutritive value for ruminants of cell mass from lysine production (CMLP) which is a by-product of the lysine manufacturing process. Proximate analysis, protein fractionation, and in vitro protein degradation using protease from Streptomyces griseus and strained ruminal fluid were carried out to estimate ruminal protein degradability of CMLP with two reference feedstuffs-soybean meal (SBM) and fish meal (FM). Amino acid composition and pepsin-HCl degradability were also determined to evaluate postruminal availability. CMLP contained 67.8% crude protein with a major portion being soluble form (45.4% CP) which was composed of mainly ammonium nitrogen (81.8% soluble CP). The amount of nucleic acids was low (1.15% DM). The total amount of amino acids contained in CMLP was 40.60% DM, which was lower than SBM (47.69% DM) or FM (54.08% DM). CMLP was composed of mainly fraction A and fraction B2, while the protein fraction in SBM was mostly B2 and FM contained high proportions of B2 and B3 fractions. The proportion of B3 fraction, slowly degradable protein, in CP was the highest in fish meal (23.34%), followed by CMLP (7.68%) and SBM (1.46%). CMLP was degraded up to 51.40% at 18 h of incubation with Streptomyces protease, which was low compared to FM (55.23%) and SBM (83.01%). This may be due to the insoluble portion of CMLP protein being hardly degradable by the protease. The in vitro fermentation by strained ruminal fluid showed that the amount of soluble fraction was larger in CMLP (40.6%) than in SBM (17.8%). However, because the degradation rate constant of the potentially degradable fraction of CMLP (2.0%/h) was lower than that of SBM (5.8%/h), the effective ruminal protein degradability of CMLP (46.95%) was slightly lower than SBM (53.77%). Unavailable fraction in the rumen was higher in CMLP (34.0%) compared to SBM (8.8%). In vitro CP degradability of CMLP by pepsin was 80.37%, which was lower than SBM (94.42%) and FM (89.04%). The evaluation of protein degradability using different approaches indicated that soluble protein in CMLP may supply a large amount of ammonia in the rumen while insoluble protein can be by-passed from microbial attacks due to its low degradability. The results from this study suggest that CMLP can be used as a protein supplement to ruminants for supplying both non-protein nitrogen to rumen microbes and rumen undegradable protein to the host animal.

Distribution of Insulin-Like Growth Factor-Binding Proteins(IGFBPs) and IGFBP-3 Proteolysis in Noninsulin-Dependent Diabetes Mellitus Serum (인슐린 비의존형 당뇨병 환자의 혈청 중 Insulin-Like Growth Factor-Binding Proteins(IGFBPs)의 분포 및 IGFBP-3의 분해)

  • Lee, Hwa-Jin;Kim, Sung-Hyun;Kwon, Mi-Jin;Nam, Taek-Jeong
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.26 no.2
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    • pp.285-290
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    • 1997
  • The insulin-like growth factors(IGFs) are bound to several binding proteins(IGFBPs) that appear to regulate IGF transport, receptor binding, and its action. The concentration of these peptides are altered by catabolic conditions. To determine IGF-I and IGFBP levels in noninsulin-dependent diabetes mellitus (NIDDM), sera was obtained from 5 patients and 7 controls. Serum levels of IGF-I in NIDDM were lower than those in either of the controls. By western immunoblot analysis, especially IGFBP-1 levels are increased, whereas IGFBP-3 levels decreased and their fragments was increased in NIDDM serum. IGFBP-3 proteolytic activity in NIDDM sera was inhibited by phenylmethylsulfonylfluoride (PMSF), aprotinin, and ethylenediaminetetraacetic acid(EDTA). This pattern of inhibition was consistent with a metal-dependent serine protease. By gelatin zymography, these proteolytic enzymes were identified as the size of 97 and 69 kDa. IGFBP-1, which is primarily insulin regulated, was increased in NIDDM and may modulate circulating IGF-I levels by regulating capillary passage of IGF-I. IGFBP-3 proteolysis markedly reduces its affinity for the IGFs, particularly for IGF-I. This accelerates their kinetics of dissociation, thereby increasing the proportions of IGF-I in free form and its availability to the cells.

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Microbiological and Enzymatic Characteristics in Alaska Pollack Sikhae during Fermentation (전통 명태식해 숙성중의 미생물 및 효소학적 특성)

  • Cha, Yong-Jun;Kim, So-Jung;Jeong, Eun-Jeong;Kim, Hun;Choi, Woo-Jin
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.33 no.10
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    • pp.1709-1714
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    • 2004
  • The changes of microflora and enzyme activities in Alaska pollack sikhae were evaluated in 3 different temperature conditions, 5$^{\circ}C$, 2$0^{\circ}C$ and alternating temperature (stored at 5$^{\circ}C$ after 10 days of fermentation at 2$0^{\circ}C$), respectively. The number of proteolytic bacteria and 2 lactic acid bacteria including Lactobacillus sp. and Pediococcus sp. increased rapidly up to 10 days and composed major portion of total viable cell (TVC) in sikhae fermented at 2$0^{\circ}C$, whereas those of TVC were occupied by Lactobacillus sp., Pediococcus sp. and yeast after 10 days of fermentation. The major species of microflora in sikhae fermented at alternating temperature were, composed of Lactobacillus, Pediococcus and Streptococcus after 10 days of fermentation. Especially, Leuconostoc sp. was kept up to 27 days at 5$^{\circ}C$ than other temperature conditions (16 days). The activities of protease and lipase in acidic region (pH 3.0) were higher at 2$0^{\circ}C$ than at 5$^{\circ}C$ due to sensitivity of temperature, although those of protease and lipase in neutral region (pH 7.0) were not found any differences in both temperatures. Changing temperature condition from 2$0^{\circ}C$ to 5$^{\circ}C$ in alternating temperature inactivated protease activity, whereas lipase activity was still maintained during fermentation.

Bitter Peptides Derived from ${\alpha}_{s1}-and\;{\beta}-Casein$ Digested with Alkaline Protease from Bacillus subtilis (Bacillus subtilis의 염기성 프로테아제로 분해된 ${\alpha}_{s1}$- 및 ${\beta}$-카세인에서 분리된 쓴 맛 펩타이드)

  • Sohn, Kyung-Hyun;Lee, Hyong-Joo
    • Korean Journal of Food Science and Technology
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    • v.20 no.5
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    • pp.659-665
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    • 1988
  • The ${\alpha}_{s1}$-and ${\beta}$-casein were purified by DEAE-cellulose chromatography and digested with alkaline protease from Bacillus subtilis. Bitter fractions from the hydrolyzates were isolated using n-butanol extraction, Sephadex G-25 gel chromatography, and high performance liquid chromatography. Peptide mixtures were separated by reverse-phase octadecyl silica column with linear gradient of 0-80% acetonitrile containing 0.1% trifluoroacetic acid. Major peaks were combined from replicate chromatographies and the bitterness of each peak was evaluated. The bitter-tasting peaks were rechromatograpied until isolated peaks were obtained. Three different bitter peptides(BP-I, BP-II, BP-III) were obtained from the ${\alpha}_{s1}$-casein hydrolyzate. BP-I was eluted at 34% acetonitrile and BP-II, 35%, BP-III, 26%, respectively. Two bitter peptides(BP-IV, BP-V) were isolated from the ${\beta}-casein$ hydrolyzate: BP-IV was eluted at 40% acetonitrile and BP-V, 42%. BP-V was the most hydrophobic peptide in the five bitter peptides. However, BP-I and BP-II tasted more bitter than BP-IV and BP-V.

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Molecular Cloning and Expression of a Novel Protease-resistant GH-36 $\alpha$-Galactosidase from Rhizopus sp. F78 ACCC 30795

  • Yanan, Cao;Wang, Yaru;Luo, Huiying;Shi, Pengjun;Meng, Kun;Zhou, Zhigang;Zhang, Zhifang;Yao, Bin
    • Journal of Microbiology and Biotechnology
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    • v.19 no.11
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    • pp.1295-1300
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    • 2009
  • A 2,172-bp full-length gene (aga-F78), encoding a protease-resistant $\alpha$-galactosidase, was cloned from Rhizopus sp. F78 and expressed in Escherichia coli. The deduced amino acid sequence shared highest identity (45.0%) with an $\alpha$-galactosidase of glycoside hydrolase family 36 from Absidia corymbifera. After one-step purification with a Ni-NTA chelating column, the recombinant Aga-F78 migrated as a single band of ~82 and ~210 kDa on SDS-PAGE and nondenaturing gradient PAGE, respectively, indicating that the native structure of the recombinant Aga-F78 was a trimer. Exhibiting the similar properties as the authentic protein, purified recombinant Aga-F78 was optimally active at $50^{\circ}C$ and pH 4.8, highly pH stable over the pH range 5.0-10.0, more resistant to some cations and proteases, and had wide substrate specificity (pNPG, melidiose, raffinose, and stachyose). The recombinant enzyme also showed good hydrolytic ability to soybean meal, releasing galactose of $415.58\;{\mu}g/g$ soybean meal. When combined with trypsin, the enzyme retained over 90% degradability to soybean meal. These favorable properties make Aga-F78 a potential candidate for applications in the food and feed industries.

Production and Characterization of Ethanol- and Protease-Tolerant and Xylooligosaccharides-Producing Endoxylanase from Humicola sp. Ly01

  • Zhou, Junpei;Wu, Qian;Zhang, Rui;Yang, Yuying;Tang, Xianghua;Li, Junjun;Ding, Junmei;Dong, Yanyan;Huang, Zunxi
    • Journal of Microbiology and Biotechnology
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    • v.23 no.6
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    • pp.794-801
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    • 2013
  • This paper reports the production and characterization of crude xylanase from the newly isolated Humicola sp. Ly01. The highest (41.8 U/ml) production of the crude xylanase was obtained under the optimized conditions (w/v): 0.5% wheat bran, 0.2% $KH_2PO_4$, and 0.5% peptone; initial pH 7.0; incubation time 72 h; $30^{\circ}C$; and 150 rpm. A considerable amount of the crude xylanase was induced using hulless barley bran or soybean meal as the carbon source, but a small amount of the enzyme was produced when supplementary urea was used as the nitrogen source to wheat bran. The crude xylanase showed apparent optimal cellulase-free xylanase activity at $60^{\circ}C$ and pH 6.0, more than 71.8% of the maximum xylanase activity in 3.0-30.0% (v/v) ethanol and more than 82.3% of the initial xylanase activity after incubation in 3.0-30.0% (v/v) ethanol at $30^{\circ}C$ for 2 h. The crude xylanase was moderately resistant to both acid and neutral protease digestion, and released 7.9 and 10.9 ${\mu}mol/ml$ reducing sugar from xylan in the simulated gastric and intestinal fluids, respectively. The xylooligosaccharides were the main products of the hydrolysis of xylan by the crude xylanase. These properties suggested the potential of the crude enzyme for being applied in the animal feed industry, xylooligosaccharides production, and high-alcohol conditions such as ethanol production and brewing.

Synthesis and Functional Properties of Plastein from the Enzymatic Hydrolysates of Filefish Protein. 3. Functional Properties of Plasteins (말쥐치육 단백질의 효소적 가수분해물을 이용한 Plastein의 합성 및 그 물성 , 3. Plastein의 기능성)

  • KIM Se-Kwon;LEE Eung-Ho
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.20 no.6
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    • pp.582-590
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    • 1987
  • Plasteins were synthesized from a peptic filefish protein hydrolysate by papain, $\alpha-chymotrypsin$ and protease(from Streptomyces griceus) under the optimum conditions of previous paper. L-glutamic acid diethylester and L-leucine ethylester were incorporated into plastein during the plastein reaction by papain. The structural changes of freeze-dried filefish meat, peptic hydrolysate, FPC and plasteins were observed by Scanning Electron Microscopy(SEM). The functional properties of plasteins also were measured. The solubility of plasteins was higher than that of FPC and the Glu-plastein had $95\%$ solubility in the range of pH 3-10. The dispersibility of Glu-plastein and protease plastein was similar to that of egg albumin, but those of the other plasteins were lower. The water holding capacity of plasteins was lower than that of egg albumin and C. Lipid absorption of Leu-plastein was tile highest, holding 1.80 ml/g, and that of the other plasteins was similar to that of egg albumin. The emulsifying activity of Leu-plastein was the highest, holding $61.2\%$, and that of Glu-plastein was the lowest, holding $50.7\%$. The emulsifying stability of plasteins was similar to that of the emulsifying activity. The emulsifying capacity of Leu-plastein was 384 ml/g(the highest), but that of Glu-plastein and $\alpha-chymotrypsin$ plastein was 248 ml/g(the lowest). The Leu-plastein shelved the highest foaming capacity, $373\%$. The foaming capacity of other plasteins was higher than that of egg albumin. The foaming stability of plasteins was superior to that of egg albumin. The viscosity of plasteins was lower than that of egg albumin. The microstructure of $\alpha-chymotrypsin$ plastein by SEM wassimilar to that of papain plastein, but other plasteins showed differences in their microstructure. The microstructure of Glu-plastein had a smooth shape.

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Quality characteristics of Doenjang prepared with sweet potato (고구마를 이용한 된장의 품질 특성)

  • Cha, Su-Jin;Park, Sao-Ra;Kim, Dong-Han
    • Food Science and Preservation
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    • v.24 no.2
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    • pp.221-229
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    • 2017
  • The effect of sweet potato on the quality of Doenjang was investigated during fermentation. Viable cells of yeast decreased gradually after 4 weeks of fermentation, but those of aerobic bacteria increased in the late stage. Amylase activity of Doenjang was higher in the late stage of fermentation, while neutral protease maintained high activity during fermentation. Hunter L and b values of Doenjang decreased gradually during fermentation, while a value was increased. The pH of Doenjang decreased gradually until 10 weeks of fermentation, and the titratable acidity was low in the sweet potato added groups. The acid value was low in the Shinyulmi sweet potato added Doenjang. Water activity and oxidation-reduction potential of Doenjang decreased during fermentation. Reducing sugar of Doenjang decreased in the middle stage of fermentation, and it was low in sweet potato added groups. The alcohol content of Doenjang decreased after 2 weeks of fermentation. Amino and ammonia-type nitrogen of Doenjang increased during fermentation and reached the maximum after 10 and 12 weeks of fermentation, respectively. After 12 weeks fermentation, 8% of Shinyulmi sweet potato added Doenjang was more favorable taste, flavor and overall acceptability (p<0.05) than the control or the Yeonwhangmi sweet potato added groups.

Effect of the Extracts from Fermented-Rhus verniciflua Stem Bark with Fomitella fraxinea on the Growth and Enzyme Activity of Soybean Product-fermenting Microorganisms (장수버섯 배양으로 제조한 발효옻 추출물이 장류 미생물의 증식 및 효소활성에 미치는 영향)

  • Choi, Han-Seok;Yeo, Soo-Hwan;Jeong, Seok-Tae;Choi, Ji-Ho;Kang, Ji-Eun;Kim, Myung-Kon
    • The Korean Journal of Mycology
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    • v.40 no.4
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    • pp.235-243
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    • 2012
  • We studied the effect of fermented Rhus verniciflua stem bark (FRVSB) extract (used in herbal med-icine by Koreans) on the microbial growth and enzyme activity of 12 soybean-fermenting microorganisms, including Bacillus spp., lactic acid bacteria, yeast, and other harmful bacteria. The ethanol and methanol extracts of FRVSB inhibited the growth of Bacillus subtilis, Bacillus licheniformis, Bacillus cereus, and Zygosaccharomyces rouxii, and in the disk diffusion assay, their inhibition zone diameters were 11.06-12.23, 12.32-18.38, 11.47-11.84, and 13.59-14.21 mm, respectively. The water extract did not show any inhibitory effect. In fact, the water extract addition enhanced the growth of B. subtilis and B. licheniformis by 1.3-4.5 fold and that of B. cereus by 1.2-1.4 fold. However, the water extract did not affect the growth of Lactobacillus plantarum, Lactobacillus mesenteroides, Saccharomyces cer-evisiae, and Escherichia coli. The addition of water extract increased the amylase and protease activity of B. subtilis and B. licheniformis.

Isolation and Characteristics of Bacteria Showing Biocontrol and Biofertilizing Activities (생물방제 및 생물비료 활성을 가지는 세균의 분리 및 특성)

  • Jung, Ho-Il;Kim, Keun-Ki;Park, Hyean-Cheal;Lee, Sang-Mong;Kim, Yong-Gyun;Kim, Hong-Sung;Lee, Cnung-Yeol;Son, Hong-Joo
    • Journal of Life Science
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    • v.17 no.12
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    • pp.1682-1688
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    • 2007
  • To develop multifunctional microbial inoculant, microorganisms with antagonistic activity and biofertilizing activity were screened. Pantoea agglomerans and Bacillus megaterium from our laboratory culture collection, and strain MF12 from soil near poultry farm in Miryang were selected. On the basis of morphological, physiological studies and 16S rDNA sequence analysis, isolate MF12 was identified as the Bacillus pumilis. Three strains were studied for insoluble phosphate solubilization, indole-3-acetic acid (IAA) and siderophore production, ammonification ability, hydrolytic enzyme production and antifungal activity against phytopathogenic fungi. P. agglomerans did not produce any visible clear zone on agar plate containing 0.5% $Ca_3(PO_4){_2}$ as a sole phosphorus source. However, this strain could solubilize insoluble phosphate in liquid medium. All strains produced IAA ranged from $3{\sim}639{\mu}g/ml$ depending on culture time and had ammonification ability. Among three strains, only P. agglomerans produced siderophore. P. agglomerans produced pectinase and lipase, B. megaterium produced amylase, protease and lipase while B. pumilis produced protease and lipase. P. agglomerans showed antifungal activities against phytopathogenic fungi, Fusarium oxysporum and Colletotrichum gloeosporioides. B. pumilis showed antifungal activities against Botrytis cinerea, Sclerotinia sclerotiorum and Phythium ultimum.