灰树花深层发酵菌丝体多糖的酶法提取及其抗肿瘤作用

对灰树花深层发酵菌丝体多糖的酶法提取工艺及其抗肿瘤作用进行了研究 .结果表明 :利用复合酶法处理并结合热水浸提 ,缩短了提取时间 ,且能显著提高多糖的提取率 ,多糖提取率达7.4 % ,效果优于其他浸提方法 .动物体内实验证明 ,腹腔注射菌丝体多糖对小鼠移植性肿瘤S180有明显的抑制作用 ,抑制率达 57.6 %～ 6 6 .7% .     

Prediction and measurement of surfactant action upon human skin under realistic conditions

The lysosomal enzyme acid phosphatase has been characterized and quantified in tapestrip biopsies of human stratum corneum by means of a sensitive spectrofluorometric procedure. When the stratum corneum of panellists was exposed to dilute solutions of various surfactants under realistic exposure conditions, the changes observed in stratum corneum acid phosphatase specific activity have been found to correlate very closely with the visual, macroscopic changes such as dryness and flakiness, that are elicited in skin as a result of surfactants. This method monitors denaturation of stratum corneum proteins, which is an important feature of skin surfactant interactions, and serves as an effective, non-invasive predictive tool for skin irritancy and mildness of surfactants.
Prevention et mesure de l'action des tensio-actifs sur la peau humaine dans des conditions conformes a la réalité     

生物工程学和发酵技术

生物工程是应用生物有机体,生物系统或生物过程来制造工业产品的学科。生物工程在应用微生物方面有很悠久的历史。例如酿酒,发酵乳制品奶酪、酸乳等。目前又在发酵罐设计,发酵工艺,酶技术及遗传工程方面具有令人振奋的进展。生物工程学着重微生物学,生物化学和化学工程的结合,并直接付之实用。本综述主要介绍生物工程中几个同发酵有关的内容,即:微生物技术、酶技术及发酵枝术。     

山奈酚抑制α-淀粉酶作用的研究

采用酶动力学方法和荧光光谱法研究山奈酚对α-淀粉酶的抑制作用。在pH6.8、37℃条件下反应20min,山奈酚（1mg/mL,0.05mL）对α-淀粉酶（0.328U/mL,0.2mL）催化活性的抑制率达到24.39%。该抑制过程是以非竞争性方式进行。同时,山奈酚对α-淀粉酶的内源荧光产生有规律的猝灭作用,以静态猝灭方式为主。二者自发结合形成复合物,其主要作用力为疏水作用,有1个结合位点。      

黄腐酸对小鼠肝酶活性的影响

观察小鼠饲料添加黄腐酸饲养19周后肝酶组织的化学变化。结果显示:试验组与对照组相比,肝细胞内琥珀酸脱氢酶、单胺氧化酶和肝巨噬细胞内酸性磷酸酶的活性均有不同程度的增强。表明黄腐酸在促进肝细胞的物质代谢和增强肝生物转化功能等方面有一定的作用。     

Amphiphilic Pentablock Copolymers Prepared from Pluronic and ε-Caprolactone by Enzymatic Ring Opening Polymerization

Amphiphilic copolymers are appealing materials because of their interesting architecture and tunable properties. In view of their application in the biomedical field, the preparation of these materials should avoid the use of toxic compounds as catalysts. Therefore, enzymatic catalysis is a suitable alternative to common synthetic routes. Pentablock copolymers (CUC) were synthesized with high yields by ring-opening polymerization of ε-caprolactone (ε-CL) initiated by Pluronic (EPE) and catalyzed by Candida antarctica lipase B enzyme. The variables to study the structure–property relationship were EPEs’ molecular weight and molar ratios between ε-CL monomer and EPE macro-initiator (M/In). The obtained copolymers were chemically characterized, the molecular weight determined, and morphologies evaluated. The results suggest an interaction between the reaction time and M/In variables. There was a correlation between the differential scanning calorimetry data with those of X-ray diffraction (WAXD). The length of the central block of CUC copolymers may have an important role in the crystal formation. WAXD analyses indicated that a micro-phase separation takes place in all the prepared copolymers. Preliminary cytotoxicity experiments on the extracts of the polymer confirmed that these materials are nontoxic.     

Keratanase II-catalyzed synthesis of keratan sulfate oligomers by using sugar oxazolines as transition-state analogue substrate monomers: a novel insight into the enzymatic catalysis mechanism

Keratan sulfate (KS) oligomers with well-defined structures were synthesized by keratanase II (KSase II)-catalyzed transglycosylation. N-Acetyllactosamine [Galbeta(1-->4)GlcNAc; LacNAc] oxazoline derivatives with sulfate groups at the C-6 (1 a) and both the C-6 and the C-6' (1 b) were prepared as transition-state analogue substrate monomers for KSase II. Monomer 1 a was effectively oligomerized by the enzyme under weak alkaline conditions, to give alternating 6-sulfated KS oligomers (2 a) in good yields, and with total control of regioselectivity and stereochemistry. KSase II also recognized 1 b, which provided fully 6-sulfated KS oligomers (2 b) in good yields under similar conditions. Nonsulfated LacNAc oxazoline was difficult to oligomerize enzymatically. These results imply that the catalysis mechanism of KSase II involves a sugar oxazolinium ion that requires the 6-sulfate group in the GlcNAc residue not only in hydrolysis of KS chains, but also in oligomerization of oxazoline monomers. This is the first report of KSase II-catalyzed transglycosylation to form beta(1-->3)-glycosidic bond through a substrate-assisted mechanism.     

A lysine to arginine substitution at position 146 of rabbit aldolase A changes the rate-determining step to Schiff base formation

Lys146 of rabbit aldolase A [D-fructose-1,6-bis(phosphate):D-glyceraldehyde-3-phosphate lyase, EC 4.1.2.13 [EC] ] was changedto arginine by site-directed mutagenesis. The k_cat of the resultingmutant protein, K146R, was 500 times slower than wild-type insteady-state kinetic assays for both cleavage and condensationof fructose-1,6-bis(phosphate), while the Km for this substratewas unchanged. Analysis of the rate of formation of catalyticintermediates showed K146R was significantly different fromthe wild-type enzyme and other enzymes mutated at this site.Single-turnover experiments using acid precipitation to trapthe Schiff base intermediate on the wild-type enzyme failedto show a build-up of this intermediate on K146R. However, K146Rretained the ability to form the Schiff base intermediate asshown by the significant amounts of Schiff base intermediatetrapped with NaBH₄. In the single-turnover experiments it appearedthat the Schiff base intermediate was converted to productsmore rapidly than it was produced. This suggested a maximalrate of Schiff base formation of 0.022 s^–1, which wasclose to the value of k_cat for this enzyme. This observationis strikingly different from the wild-type enzyme in which Schiffbase formation is >100 times faster than k_cat. For K146Rit appears that steps up to and including Schiff base formationare rate limiting for the catalytic reaction. The carbanionintermediate derived from either substrate or product, and theequilibrium concentrations of covalent enzyme-substrate intermediates,were much lower on K146R than on the wild-type enzyme. The greaterbulk of the guanidino moiety may destabilize the covalent enzyme-substrateintermediates, thereby slowing the rate of Schiff base formationsuch that it becomes rate limiting. The K146R mutant enzymeis significantly more active than other enzymes mutated at thissite, perhaps because it maintains a positively charged groupat an essential position in the active site or perhaps the Argfunctionally substitutes as a general acid/base catalyst inboth Schiff base formation and in subsequent abstraction ofthe C4-hydroxyl proton.