Autolysis and the endogenous proteinases characterised in beardless barb (Anematichthys apogon) muscle

Beardless barb is a common fish species used in fermentation of fish paste Ka-pi-plaa. Autolytic profile of beardless barb muscle showed the maximum autolysis was at 50 °C, at both acidic and alkaline pH values. With augmentation concentration of NaCl, autolytic activity slightly decreased. Endogenous proteinases isolated from fish muscle in crude extract forms were also characterised. The acidic proteinases had optimum activity at pH 3.0 and 50°C, and they showed higher proteolytic activity than the alkaline proteinases which were optimally active at pH 9.0 and 50 °C. Proteinases in peak at pH 3.0 were inhibited by pepstatin A, but those in peak at pH 9.0 were highly inhibited by PMSF, TLCK and soybean trypsin inhibitor, suggesting that both aspartic and serine proteinases were existed in beardless barb muscle. The proteinases were stable in pH range of 2.0-5.0 but unstable at the temperatures higher than 40 °C. NaCl suppressed the proteolytic activity, ATP activated the proteinase activity, while CaCl₂, MgCl₂ and CoCl₂ exhibited no influence on the activity. The results implied that cathepsin D is the predominant proteinase responsible for autolysis in beardless barb. The findings were useful to improve the processing and qualities of Ka-pi-plaa product using beardless barb as raw material.     

The Loss of Hydrophobic Polypeptides during Fermentation and Conditioning of High Gravity and Low Gravity Brewed Beer

The object of this study was to investigate the loss of hydrophobic polypeptides, which are important for foam quality and stability in finished beer. Loss of hydrophobic polypeptide due to fermenter foaming occurs during transfer of fermented wort since a gradient of hydrophobic polypeptides towards the surface is created during fermentation. Due to higher polyphenol levels in high gravity (20°Plato) wort, more hydrophobic polypeptides are lost due to cold break (cold trub) precipitation compared to low gravity (12°Plato) wort. Another important factor affecting the loss of hydrophobic polypeptides could be proteinase A activity during fermentation, especially in high gravity fermentation where the yeast is exposed the higher stress. During high gravity fermentation, where osmotic pressures are higher, ethanol levels become greater, and nitrogen‐carbohydrate ratios are lower, more proteinase A is released by the yeast. This release of proteinase A into fermenting wort could have implications for the foam stability of the finished product.     

柞蚕丝织物的柔软整理方法分析

采用蛋白酶和有机硅柔软剂对柞蚕丝织物进行柔软整理,通过正交试验,确定了两种整理的最佳工艺条件,并对比分析了两种整理方法在柔软效果、工艺条件和整理成本上的差异,得出柞蚕丝织物采用有机硅柔软整理优于蛋白酶整理的结论。     

信号肽引导源基因在昆虫表达系统中的分泌表达

将杆状病来源的gp67信号肽引导的慈姑蛋白酶抑制剂(API)基因与植物来源的慈姑蛋白酶抑制剂信号肽引导的慈姑蛋白抑制剂(API)基因，分别克隆到昆虫杆状病毒转载体pBacPAK8中，通过共转染将它们分别整合到昆虫病毒表达载体Bm—BacPAK6中，将获得的重组病毒CrBK—API2和CrBK—bpAPI4分别接种家蚕(Bombyx mori)细胞、家蚕幼虫及蛹体中，慈姑蛋白酶抑制剂得到了高效表达。从家蚕细胞的表达来看，外源表达产物大多存在于细胞外的上清液中；从家蚕幼虫体内表达来看，信号肽能引导外源基因高效表达，且表达产物的90％以上能分泌到细胞外；蛹体中的表达情况与幼虫中的相似，但表达产物出了不均一的现象，从表达产物的分子量测定来看，这两种信号肽在表达过程中都没有被切除。     

酶解乳蛋白制取乳源肽的研究

用酸性蛋白酶、中性蛋白酶和碱性蛋白酶对牛乳蛋白进行水解，研究了水解工艺，并通过正交试验对工艺进行了优化。结果为：酸性蛋白酶水解牛乳蛋白的最佳工艺条件是：水解温度50℃，时间120min，pH值3．0，加酶量8．0％；中性蛋白酶水解温度50℃，时间90min，pH值7．0，加酶量1．60％；碱性蛋白酶水解温度60℃，时间105min，pH值9．0，加酶量8．0％。中性蛋白酶水解物苦昧最小，最适宜作食品。     

变性豆粕中蛋白质的酶水解特性的研究

变性豆粕是由大豆浸油后,经高温脱溶所得。本文以高温变性脱脂大豆粕为原料,用正交实验法对变性后在蛋白酶作用下的水解特性进行了深入研究。选用国产1398中性蛋白酶为水解酶对变性豆粕进行水解,研究了变性豆粕中蛋白质溶出率随温度、pH值、时间、底物浓度及用酶量的变化规律,找到了水解变性豆粕的最佳实验条件。为生产实践提供了基础数据,该研究结果对其它蛋白质原料的水解特性研究也具有参考价值。研究结果表明：1398中性蛋白酶水解高温变性豆粕的最佳条件为：温度45℃,时间3h,底物浓度1．0％,用酶量1600u/g,pH值7．0,在此条件下,变性豆粕中蛋白质可有90．71％水解溶出。     

水解动物蛋白(HAP)的酶法制备及应用

报道了畜禽肉及鱼用酶解法制备ＨＡＰ的最适条件。当用双酶水解时，先用胰蛋白酶后用胃蛋白酶，水解效果最好。酶—酸联合水解，胶体磨处理及热变性，都能提高蛋白质水解率。ＨＡＰ有广泛应用前景。     

酶解法制备米糠膳食纤维

对米糠中膳食纤维的酶解法制备工艺进行了研究。通过单因素试验、正交试验和极差分析,得出了酶法制备米糠膳食纤维的最佳工艺参数。结果表明,在混合酶(α-淀粉酶与糖化酶质量之比为1∶2)用量0.4%、70℃处理45min,蛋白酶用量0.3%、50℃处理45min时,膳食纤维得率较高,膳食纤维中的酸性洗涤纤维含量达68.54%。因此,该酶法工艺为米糠膳食纤维的制备提供了依据。     

蛋白酶对大豆分离蛋白的降解模式研究

采用十二烷基酸钠聚丙烯酰胺凝胶电泳（SDS-PAGE）方法，分析了几种商品蛋白酶（包括枯草杆菌蛋白酶、胰蛋白酶、胰凝乳蛋白酶、木瓜蛋白酶和细菌碱性蛋白酶）对大豆分离蛋白（SPI）的降解模式。结果表明，大豆球蛋白酸性亚基的Ax多肽链最容易被水解，所有的酶对其均有作用，而碱性亚基和大豆伴球蛋白的B亚基最难被水解。木瓜蛋白酶对大豆分离蛋白的水解最迅速、彻底，由于大豆蛋白中含有胰蛋白酶抑制因子，胰蛋白酶和胰凝乳蛋白酶对大豆蛋白水解能力较差。     

High γ-aminobutyric acid production from lactic acid bacteria: Emphasis on Lactobacillus brevis as a functional dairy starter

γ-Aminobutyric acid (GABA) and GABA-rich foods have shown anti-hypertensive and anti-depressant activities as the major functions in humans and animals. Hence, high GABA-producing lactic acid bacteria (LAB) could be used as functional starters for manufacturing novel fermented dairy foods. Glutamic acid decarboxylases (GADs) from LAB are highly conserved at the species level based on the phylogenetic tree of GADs from LAB. Moreover, two functionally distinct GADs and one intact gad operon were observed in all the completely sequenced Lactobacillus brevis strains suggesting its common capability to synthesize GABA. Difficulties and strategies for the manufacture of GABA-rich fermented dairy foods have been discussed and proposed, respectively. In addition, a genetic survey on the sequenced LAB strains demonstrated the absence of cell envelope proteinases in the majority of LAB including Lb. brevis, which diminishes their cell viabilities in milk environments due to their non-proteolytic nature. Thus, several strategies have been proposed to overcome the non-proteolytic nature of Lb. brevis in order to produce GABA-rich dairy foods.