壳聚糖固定瑞士乳杆菌蛋白酶的条件研究

以壳聚糖为载体,戊二醛为交联剂,采用交联-吸附法对瑞士乳杆菌蛋白酶的固定化条件进行研究。在单因素试验基础上,以固定化酶活力为主要指标,研究凝结液、壳聚糖质量浓度、酶用量、交联时间、戊二醛质量浓度对瑞士乳杆菌蛋白酶固定化的影响。运用响应面对固定化条件进行优化,确定瑞士乳杆菌蛋白酶的最优固定条件：凝结液为4g/100mL NaOH-甲醇(体积比3:1)、壳聚糖质量浓度2.89g/100mL、酶用量2.95mg、交联时间1h、戊二醛质量浓度0.40g/100mL,此时固定化酶活力为28.67U。     

蛋白酶A对纯生啤酒泡持性的影响研究

该文研究蛋白酶A活力对纯生啤酒泡持性的影响,同时建立初始成品纯生啤酒及过滤前发酵液蛋白酶A活力限量值。结果表明,随着贮存时间的延长,纯生啤酒蛋白酶A活力和泡持值均呈下降趋势,最终残留蛋白酶A活力是影响其货架期泡持值的主要因素。纯生啤酒泡持值与蛋白酶A活力呈显著负相关（P＜0.01）。为了保证成品纯生啤酒在货架期内泡沫稳定性,发酵液出罐滤酒前的蛋白酶A活力应＜24×10-5 U/mL,相应成品纯生啤酒初始蛋白酶A活力应＜15×10-5 U/mL。该内控标准能为纯生啤酒生产企业控制泡持性提供指导。     

响应面法优化预酶解提取米渣蛋白的研究

为改善米渣中蛋白质的提取效果,采用中性蛋白酶酶解、碱溶两步法提取米蛋白.固定碱提取工艺,通过单因素实验,确定了预酶解的基本条件;并以pH和加酶量和反应时间为自变量,蛋白质提取率和质量分数为响应值,采用Box Behnken试验设计方法,对预酶解条件进行响应面（RSM）优化,确定了预酶解的最优工艺.最优条件下得到米渣蛋白质的提取率为78.27%,质量分数为72.86%,较直接碱提取,两者均有了一定的提高,且以蛋白质提取率提升幅度大,且优化条件下的实验结果与RSM回归方程预测值吻合良好.     

芽孢杆菌的产酶特性及其对抗生素的耐受性

以4株枯草芽孢杆菌（Bacillus subtilis）和2株地衣芽孢杆菌（Bacillus licheniformis）为研究对象,通过测定其成胞率、中性蛋白酶活性及对24种抗生素的耐受性,从中筛选优良芽孢杆菌。结果表明,从6株芽孢杆菌中筛选得到3株成胞率和产中性蛋白酶均较好的芽孢杆菌,分别为枯草芽孢杆菌BLCC1-0552、BLCC1-0615和地衣芽孢杆菌BLCC1-0441,液体发酵24 h后,成胞率较高,均达到98%;中性蛋白酶活性分别为90.58 U/mL、100.32 U/mL和24.72 U/mL。其中,枯草芽孢杆菌BLCC1-0615固体发酵玉米-豆粕型常规饲料产中性蛋白酶活力最高,发酵48 h时达到2 154.49 U/g。3株芽孢杆菌对抗生素的耐受性不一致,其中枯草芽孢杆菌BLCC1-0615对24种抗生素中的17种表现敏感,敏感率最高为70.83%。因此,确定优良菌株为枯草芽孢杆菌BLCC1-0615,其成胞率、产中性蛋白酶能力好及对抗生素耐受性较低,具有作为饲料添加剂应用于畜禽饲料中的潜力。     

Degradation of myofibrillar proteins by a myofibril-bound serine proteinase in the skeletal muscle of crucian carp (Carasius auratus)

A myofibril-bound serine proteinase (MBSP) in the skeletal muscle of crucian carp (Carasius auratus) was identified. Hydrolysis of myofibrillar proteins by the endogenous MBSP was studied. Myosin heavy chain (MHC) was degraded markedly when crucian carp myofibril was incubated at 55 °C, as shown by SDS-PAGE. Prolonged incubation of myofibril at 55 °C also caused the obvious degradation of tropomyosin, while the decomposition of other myofibrillar proteins, such as α-actinin and actin, was slight, as detected by Western blotting. The results suggest the existence of an endogenous myofibril-associated proteinase in crucian carp myofibril, which efficiently cleaves MHC and tropomyosin. Serine proteinase inhibitors (Lima bean trypsin inhibitor, PMSF and benzamidine) greatly suppressed the degradation of MHC, caused by the enzyme, while inhibitors for cysteine, metallo-, and aspartic proteinases showed only partial or incomplete inhibitory effects, indicating that the endogenous proteinase is a serine proteinase. Substrate specificity analysis, using partially purified crucian carp MBSP, suggested that the enzyme is a trypsin-like serine proteinase.     

酶制剂在大豆蛋白水解中的应用

制剂在食品加工领域中应用广泛，如用于植物蛋白的水解。对于大分子蛋白质，酶法改性具有理化改性无可比拟的优点：营养价值降低不显著、无不良副反应、反应条件温和、设备要求不高、作用具有独特专一性、效率高、反应进程容易控制等。本文综述了目前在大豆蛋白酶法水解领域中常用的几种酶（包括植物蛋白酶、动物蛋白酶和微生物蛋白酶）的特点及应用。     

Original features of cell-envelope proteinases of Lactobacillus helveticus. A review

Lactobacillus helveticus is a lactic acid bacterium very used in fermented milks and cheese. The rapid growth of L. helveticus in milk is supported by an efficient cell envelope proteinase (CEP) activity, due to subtilisin-like serine proteases. These enzymes play also crucial roles in texture and flavor formation in dairy products as well as in generating in situ bioactive peptides. In L. helveticus, several genes encoding putative CEPs were detected and characterized by a large intraspecific diversity; little is known about regulation of expression of CEP-encoding genes. Anchored at the bacterial surface, CEPs are large-sized enzymes (> 150 kDa) hydrolyzing β- and α_s1-casein as well. Substrate cleavages occur after almost all types of amino acids residues, but mass spectrometry analysis revealed L. helveticus strains with specific profiles of substrate hydrolysis, which could explain identification of strains associated with interesting technological properties. In this review, the most recent data regarding CEP-encoding genes, CEP activities toward caseins and L. helveticus strain diversity are discussed.     

牡蛎肉双酶复合水解工艺

以牡蛎为原料,利用多种蛋白酶对牡蛎蛋白质进行水解,挑选中性蛋白酶1398和Flavourzyme蛋白酶进行复合酶解,得出酶解的最佳工艺参数为:时间16h,中性蛋白酶1398与Flavourzyme作用的时间比为16∶15,中性蛋白酶1398酶与底物比为300U/g,Flavourzyme酶与底物比为1200U/g.水解后,氨基态氮质量分数为0.469%,总氮回收率为91%.     

蛋白酶水解牛乳蛋白的研究

探讨了几种不同的蛋白酶对牛乳蛋白的最佳酶解条件。通过对蛋白酶水解度和苦味值的比较，确定了胰蛋白酶和复合风味蛋白酶作为复配酶。其最佳酶解参数为：酶解温度55℃、时间4h、加酶量6g(每千克蛋白质中)、复合风味蛋白酶与胰蛋白酶的质量比为2：1，水解度可达25％以上，苦味值较小，满足了蛋白酶水解改性的要求。     

肉桂色曲霉9824饲用酸性蛋白酶的产生、性质及应用研究

本文对肉桂色曲霉9824菌株酸性蛋白酶产酶条件，酶学性质和在断奶仔猪的喂养效果进行了研究。结果表明，采用麸皮与豆饼比例6：4，氯化铵浓度1．5％，物料加水比1：1，菌株接种量0．3％，经66h发酵，其产酶活较高，该酶最适酸碱度pH3.0，最适作用温度为40℃，用pH4．0的乳酸缓冲液在50℃下提酶较完全。该酶具有较好的热稳定性，在100℃，1h可保持80％的酶活力。小鼠急性毒性试验证明该酶安全无毒。以0．1％添加到当地饲喂秧藤粉的断奶仔猪日粮中，经40d喂养，试验组比对照组日增重和饲料转化率分别提高13．2％和2．4％，并有效地控制了腹泻。