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Chinese Journal of Oceanology and Limnology 2009年第4期27卷 753-761页

作者：李静池振明王祥红彭莹池哲 UNESCO Chinese Center of Marine Biotechnology Ocean University of China

A total of 400 yeast strains from seawater, sediments, saltern mud, marine fish guts, and marine algae were obtained. The protease activity of the yeast cultures was estimated, after which four strains （HN3.11, Nllb, YF04C and HN4.9） capable of secreting extracellular alkaline protease were isolated. The isolated strains were identified as Aureobasidium pullulans, Yarrowia lipolytica, lssatchenkia orientalis and Cryptococcus cf. aureus. The optimal pH of the protease activity produced by strains HN3.11, YF04C, and HN4.9 was 9.0, while that of the protease produced by strain N1 lb was 10.0. The optimal temperature for protease activity was 45℃for strains HN3.11, N11b, and YF04C, and 50℃ for strain HN4.9. After digestion of shrimp （Penaeus vannamei） protein and spirulina （Arthospira platens＆） protein with the four crude alkaline proteases, the filtrate from spirulina （Arthrospira platensis） powder digested by the crude alkaline protease of strain HNYl 1 was found to have the highest antioxidant activity （61.4%） and the highest angiotensin I converting enzyme （ACE）-inhibitory activities （68.4%）. The other filtrates had much lower antioxidant activity and ACE-inhibitory activities.

关键词： marine yeasts alkaline protease yeast diversity phylogenetic analysis bioactive peptide

Regioselective Synthesis of Polymerizable Vinyl Guaifenesin Esters Catalyzed by an alkaline protease of Bacillus subtilis

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Chinese Chemical Letters 2004年第2期15卷 175-178页

作者： NaWANG QiWU JianMingXU XiuMingJIANG XianFuLIN DepartmentofChemistry ZhejiangUniversityHangzhou310027

Three polymerizable vinyl guaifenesin esters with different acyl donor carbon chain lengths (C4,C6,C10) were regioselectivly synthesized by an alkaline protease from Bacillus subtilis in pyridine at 50C for 1, 3, 5 da... 详细信息

关键词： Vinyl guaifenesin ester regioselectivity alkaline protease transesterification.

Regiospecific Addition of Uracil to Acrylates Catalyzed by alkaline protease from Bacillus subtilis

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Chinese Chemical Letters 2004年第5期15卷 594-596页

作者： YingCAI JianYiWU NaWANG XiaoFengSUN XianFuLIN DepartmentofChemistry ZhejiangUniversityHangzhou310027 DepartmentofChemistry .JiaxingCollegeJiaxing314001

Michael addition reactions of uracil to acrylates were catalyzed by an alkaline protease from Bacillus subtilis in dimethyl sulfoxide at 55 ℃ for 72 h. The adducts were determined by TLC, IR and 1H NMR.

关键词： alkaline protease Michael addition uracil pyrimidine.

Enzymatic kinetics ofβ-conglycinin using alkaline protease from Bacillus subtilis ACCC 01746 and analysis of antigenicity of hydrolyzed peptide

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Grain & Oil Science and Technology 2021年第3期4卷 141-148页

作者： Haicheng Yin Xinrui Zhang Zhixiang Yang Jin Huang College of Biological Engineering Henan University of TechnologyZhengzhou 450001China

β-Conglycinin,the main protein of soybean,is a key allergen that causes soybean allergies,and hydrolysis is usually applied to lower its *** evaluated the enzymolysis characters ofβ-conglycinin from the perspective of enzymolysis kinetics using alkaline protease from *** ACCC 01746.A dynamic model describing the hydrolysis ofβ-conglycinin was proposed using the initial substrate concentration,enzyme dosage(enzyme to substrate ratio)and hydrolysis time as variables to illustrate the kinetic behavior of enzymatic *** hydrolysis of soybeanβ-conglycinin was carried out at 60 g/L protein concentration,0.6%enzyme dosage,55℃ and pH 8.5 to observe the peptides with anti-enzymatic *** hydrolysates were gradually fractionated by ultrafiltration through cut-off membranes with molecular weights of 40,30,20,and 10 kDa,and their antigenicities were evaluated using indirect competitive enzyme-linked immunosorbent *** results showed that the degree of hydrolysis(DH)ofβ-conglycinin decreased as theβ-conglycinin concentration(S0)increased,but increased with enzyme dosage(E0)***,the enzymatic hydrolysis ofβ-conglycinin followed the first-order kinetics *** hydrolysis rate(V)was(527.89C_(E0)-2.5533C_(S0))exp(-0.022DH),the DH-hydrolysis time was 45.454ln[1+(11.614C_(E0)/C_(S0)-0.0562)t],and the correlated kinetic constants k2 and kd were 527.89 min^(−1)and 8.6126 min^(−1),*** hydrolysis behavior ofβ-conglycinin varied considerably among theα',α,andβ*** hydrolysis rates were observed for theα'andαsubunits compared to theβ*** relative molecular weights of the intercepted peptides from the hydrolysates were 14.8-40.1 kDa,and the antigenicity of the peptides with smaller molecular weight was reduced,but not removed ***,the alkaline protease from the strain appeared to effectively reduce the allergenicity ofβ-***,it is possible to produce less allergenic soybean

关键词： B.subtilis ACCC 01746 alkaline protease β-Conglycinin Enzymatic kinetics Antigenicity

Study on enzymatic hydrolysis of soybean β-conglycinin using alkaline protease from Bacillus subtilis ACCC 01746 and antigenicity of its hydrolysates

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Grain & Oil Science and Technology 2021年第1期4卷 18-25页

作者： Haicheng Yin Xinrui Zhang Jin Huang College of Biological Engineering Henan University of TechnologyZhengzhou 450001China

Due to its beneficial health effects,the use of soybean protein has shown a continuous increase,but concerns regarding the allergenicity of soybean antigenic protein have also *** study aimed to evaluate the hydrolytic effects of a non-commercial alkaline protease isolated from the Bacillus subtilis ACCC 01746 on soybeanβ-conglycinin and the allergenicity of its *** protease of the strain was separated by precipitation method of organic solvents,and theβ-conglycinin was separated by alkali-solution and acid-isolation and purified by use of gel *** the degree of hydrolysis(DH)and inhibition rate as evaluation indexes,the enzymatic hydrolysis parameters ofβ-conglycinin was optimized by single factor and L_(9)(3^(4))orthogonal tests,so as to explore the effect of the protease on the hydrolysis degree and the antigenicity ofβ-conglycinin *** results showed that the native enzyme existed as an 18.3 kDa monomer with a 430 U/g maximum *** purity ofβ-conglycinin was 84.8%.The single-factor test results showed that DH showed the oppostie trendency with the inhibition rate,and the increase of protein concentration causedmonotone increasing and monotone decreasing of the inhibition rate and the DH,and the optimal protein concentration was 30 mg/*** optimization results showed that pH had the largest impacts on both DH and the inhibition rate,followed by enzyme dosage,hydrolysis temperature and hydrolysis *** the optimum hydrolysis conditions of protein concentration 30mg/mL,enzymedosage0.7%,hydrolysis time40min,temperature 55°C and pH8.5,the DH reached the highest of 76.28%,and the inhibition rate was the lowest of 27.03%,which was reduced greatly compared with that before *** results suggested that alkaline protease appeared to show a relatively high effeciency in lowering soybean allergenicity,making it possible to produce low-allergenicity soybean protein.

关键词： alkaline protease Bacillus subtilis ACCC 01746 β-conglycinin Hydrolysis degree Antigenicity

Levels of antioxidant enzymes and alkaline protease from pulp and peel of sunflower

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Asian Pacific Journal of Tropical Biomedicine 2017年第6期7卷 533-537页

作者： Wesen Adel Mehdi Faridah Yusof Layla O.Farhan Atheer Awad Mehde Raha Ahmed Raus Department of Chemistry College of Sciences for WomenUniversity of Baghdad Department of Biotechnology Engineering College of EngineeringInternational Islamic University Malaysia Department of Technical Medical Analysis College of Health and Medical TechnologyMiddle Technical University

Objective:The activity of enzymes participating in the systems of antioxidant protection was assayed in the peel and pulp of *** essential roles of proteases in food stimulate research to find other sources of the enzyme especially from non-conventional *** the present work,we study several biochemical parameters in the pulp and peel of ***:Pulp and peel of sunflower was extracted,antioxidant enzymes and nonenzymatic antioxidant were *** protease was measured and purified from pulp in ***:High carbohydrate concentration,beta-carotene,catalase and ascorbate peroxidase activities,free radical scavenging capacity and free flavonoid content were observed in the peel of ***,MDA and ceruloplasmin activities were high in the pulp of ***:The present study concluded that peel in sunflower are strong radical scavengers and can be considered as good sources of natural antioxidants for medicinal and commercial *** analysis showed that protease activity was a significantly high in the pulp compared to the peel.

关键词： Pulp sunflower Antioxidant enzymes Free radical scavenging capacity Free flavonoid content alkaline protease

alkaline protease Production by a Strain of Marine Yeasts

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Journal of Ocean University of China 2006年第3期5卷 263-268页

作者： WANG Ping CHI Zhenming MA Chunling UNESCO Chinese Center of Marine Biotechnology Ocean University of China Qingdao 266003 P. R. China

Yeast strain 10 with high yield of protease was isolated from sediments of saltern near Qingdao, China. The protease had the highest activity at pH 9.0 and 45 ℃. The optimal medium for the maximum alkaline protease production of strain 10 was 2.5 g soluble starch and 2.0 g NaNO3 in 100 mL seawater with initial pH 6.0. The optimal cultivation conditions for the maximum protease production were temperature 24.5 ℃, aeration rate 8.0 L min -1 and agitation speed 150 r min -1. Under the optimal conditions, 623.1 U mg -1 protein of alkaline protease was reached in the culture within 30 h of fermentation.

关键词： marine yeasts alkaline protease fermentation optimal conditions

Screening of Strains Producing alkaline protease from Soil and Study on the Conditions for Enzyme Production

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Meteorological and Environmental Research 2011年第8期2卷 82-85页

作者： ZHOU Yi-jun HE Wen-jie FU Sha-ou TIAN Jun-duan MA Ting-ting College of Life and Environmental Sciences Minzu University of ChinaBeijing 100081China

[Objective] The aim was to screen strains producing alkaline protease from soil and study the conditions for enzyme production.[Method] Eight strains producing alkaline protease were isolated from soil through plate isolation,and the ability of enzyme production was measured by filter paper and Folin-phenol *** strain with the strongest ability of enzyme production was screened as a candidate strain,then the factors influencing the ability of enzyme production was studied,finally the conditions for enzyme production was optimized through orthogonal test.[Result] No.5 strain was screened as a candidate strain due to its strong ability of enzyme production(6.00 U/ml),which accounted for 134.1% of that of Bacillus licheniformis,and it was gram-positive bacterium belonging to *** test showed that the optimal condition for producing protease was an environment with pH=11,0.3% of sucrose and 0.8% of peptone in the fermentation medium,and inoculation amount was 105 cfu/*** addition,peptone had significant impact on the level of enzyme production.[Conclusion] The study could provide theoretical references for the screening of strains producing alkaline protease.

关键词： Soil Screening Strain alkaline protease China

Mutagenesis of alkaline protease-Producing Marine Strains and Enzymatic Properties

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Agricultural Biotechnology 2012年第2期1卷 32-34,37页

作者： Wei XIA Hua LI Xiaoli WANG Guoyu ZHANG Chao LI Shaohui ZHU Hao ZHANG Quanhan XU Qingdao Zhongren Pharmaceutical Co. Ltd.

[ Objective] The aim of the research was to select alkaline pmtease-producing strains and provide basis for the application in production. [ Method ] An alkaline protease-preducing strain was isolated from the sea water and sea mud collected from Qingdao coastal area. After ultraviolet mutagenesis and microwave mutagenesis, a target strain ZR-58-3-1-3 was obtained. The protease activity and enzymatic properties of the strain were determined preliminarily. [ Result ] Prote- ase activity of the selected strain reached 145 U/ml. After the compound mutagenesis, protease activity of the fermentation liquid of strain ZR-58-3-1-3 reached 775 U/m｜, which was about 4.3 times higher than that of the original strain. The optimal temperature for alkaline protease produced by strain ZR-58-3-1-3 was 45℃, and the optimal pH was 8.5, suggesting it belongs to moderate temperature protease with relatively high thermal stabihty. [ Conclusion] Strain ZR-58-3-1-3 has stable alkaline protease production performance and can be used as an alkaline protease-producing mutant strain.

关键词： alkaline protease Marine strain Mutagenesis Enzymatic properties

Production of alkaline protease by Bacillus Licheniformis

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Journal of Chemistry and Chemical Engineering 2011年第2期5卷 105-109页

作者： Bahareh Sadeghein Seyed Abolhasan Alavi Department of Chemical Engineering and Biotechnology Faculty of Engineering Science and Research Branch Islamic AzadUniversity Tehran *** Iran

In this research the results of studies on optimization of alkaline protease production by Bacillus licheniformis are reported. The parameters, which were taken into consideration, are pH, temperature, time course of enzyme production, stirring rate and kinetics parameters. The effect of various carbon and nitrogen sources in culture medium compound on enzyme production was also considered The result of optimization revealed that maximum protease production was obtained at 37 ℃, pH equivalent tol 0.0 and with 150 rpm will occur after 72 hours. By comparing the effect of 5 carbon sources （maltose, glucose, starch, casein and lactose） in enzyme production, it has been known that using lactose will increase about 1.5 times enzyme production, compared to condition in which maltose is used. The result of studies on the effect of five nitrogen sources （i.e., peptone, tryptone, ammonium sulfate, urea and corn steep liquor） shows that corn steep liqour increases enzyme production more than others, while peptone can also be considered as a good nitrogen source; but, ammonium sulfate and urea reduce enzyme production considerably. It was concluded that protease production occurs in the stationary phase of growth. Studying the kinetics parameters resulted that the best model for the enzyme above is Lineweaver-Burk model according to which Km is 0.64 mmol and Vmax is 88 lamol/min.

关键词： Bacillus licheniformis production enzyme alkaline protease