An Atomistic Understanding of Allosteric Inhibition of Glutamate Racemase: a Dampening of Native Activation Dynamics

Glutamate racemases (GR) are members of the family of bacterial enzymes known as cofactor-independent racemases and epimerases and catalyze the stereoinversion of glutamate. D-amino acids are universally important for the proper construction of viable bacterial cell walls, and thus have been repeatedly validated as attractive targets for novel antimicrobial drug design. Significant aspects of the mechanism of this challenging stereoinversion remain unknown. The current study employs a combination of MD and QM/MM computational approaches to show that the GR from H. pylori must proceed via a pre-activation step, which is dependent on the enzyme's flexibility. This mechanism is starkly different from previously proposed mechanisms. These findings have immediate pharmaceutical relevance, as the H. pylori GR enzyme is a very attractive allosteric drug target. The results presented in this study offer a distinctly novel understanding of how AstraZeneca's lead series of inhibitors cripple the H. pylori GR's native motions, via prevention of this critical chemical pre-activation step. Our experimental studies, using SPR, fluorescence and NMR WaterLOGSY, show that H. pylori GR is not inhibited by the uncompetitive mechanism originally put forward by Lundqvist et al.. The current study supports a deep connection between native enzyme motions and chemical reactivity, which has strong relevance to the field of allosteric drug discovery.     

应用高效液相色谱-电感耦合等离子体质谱联用技术分析研究畜肉中硒形态

目的 应用高效液相色谱-电感耦合等离子体质谱联用技术（HPLC-ICP/MS）分析研究畜肉中Se（Ⅵ）、Se（Ⅳ）、SeCys、MeSeCys和SeMet 5种硒形态。方法 采用超声辅助蛋白酶ⅩⅣ和胰脂肪酶酶解提取畜肉中硒形态,以C18反相色谱柱为分析柱,10 mmol/L柠檬酸和5 mmol/L己烷磺酸钠（pH=4.0,含1%甲醇）为流动相,等度洗脱,HPLC-ICP/MS分析。结果 5种硒形态可在7 min内有效分离,在线性范围内相关系数均>0.999,线性良好;Se（Ⅵ）、SeCys、MeSeCys和SeMet 4种硒形态检出限为0.000 3~0.002 mg/kg,不同浓度水平的加标回收率为80.0%~103.3%,相对标准偏差均小于6.8%,有证标准物质ERM^? BC210a测定值在标准值范围内。结论 该方法适用畜肉中Se（Ⅵ）、SeCys、MeSeCys和SeMet的分析测定。畜肉中的硒形态主要为SeMet,同时含有少量的MeSeCys和SeCys。     

固定化酶及其在化工等领域中的应用

简要介绍了固定化酶技术的概念、制备方法(包括传统固定化技术、传统固定化技术的改进方法、新型固定化技术)及其在化学化工、食品行业、临床医药、生物传感器和环境科学等领域中的应用现状与存在的问题，展望了固定化酶技术在皮革行业中的研究与应用前景。对于皮革工业的酶技术具有一定的借鉴意义和参考价值。     

嗜碱性芽孢杆菌碱性纤维素酶酶系的研究

本文研究了各种碳、氮源、ｐＨ、通风量等工艺条件对嗜碱性芽孢杆菌Ｖ－５产酶酶系的影响，通过酶在表面活性剂中的实验，分析不同表面活性剂对酶系中各组分的影响，初步研究酶系与去污力的关系，认为除以内切酶为主外，酶系中另二个组分，在去污效果中的辅助作用不容忽视。     

利用MTG酶提高精纺羊毛织物易护理性能

从理论上分析了谷氨酰胺转胺酶对羊毛纤维的作用机理,探讨了采用谷氨酰胺转胺酶处理羊毛织物的方法。对采用谷氨酰胺转胺酶处理后的精纺羊毛织物进行了测试,结果表明其尺寸稳定性、抗皱性、压烫性能等织物易护理指标均有一定程度的提高。     

Release of short and proline-rich antihypertensive peptides from casein hydrolysate with an Aspergillus oryzae protease

Angiotensin-I converting enzyme inhibitory activities were measured after hydrolysis of casein by 9 different commercially available proteolytic enzymes. Among these enzymes, a protease isolated from Aspergillus oryzae showed the highest angiotensin-I converting enzyme inhibitory activity per peptide. The A. oryzae peptide also showed the highest antihypertensive effect in spontaneously hypertensive rats when the systolic blood pressure was measured 5 h after oral administration of 32 mg/kg of various enzymatic hydrolysates. Significant antihypertensive effects were observed with dosages of 9.6, 32, and 96 mg of the A. oryzae peptide/kg of body weight (BW), and the effects were dependent on these peptide dosages.Analysis of peptide length showed the A. oryzae hydrolysate was the shortest of all tested casein hydrolysates; the peptide mixture had an average value of 1.4 amino acids (AA) in the sequence. To further characterize the A. oryzae hydrolysate, we analyzed the AA sequence of the whole peptide mixture. Various AA were detected at the first AA position, however, an increased number of Pro residues were observed at the second and third position of the A. oryzae hydrolysate. No strong signals were detected after the fourth AA position of the A. oryzae hydrolysate. These results suggest that the casein hydrolysate of A. oryzae, which expressed potent antihypertensive effects in spontaneously hypertensive rats, mainly contain short peptides of X-Pro and X-Pro-Pro sequences.     

旋光性氰醇的酶催化合成

综述了酶催化立体选择性合成及动力学折分制备旋光性氰醇的几种方法。     

桉木APMP制浆过程浆中残余木素分子量的变化

利用凝胶渗透色谱法和元素分析法测定了尾叶桉酶解木素在APMP制浆及漂白过程的分子量变化。研究结果表明：由于HOO-的特殊作用,制浆及漂白之后,浆中剩余木素的平均分子量相对有所增大。     

超声波辅助酶法处理对百香果出汁率及品质的影响

近年来,国内外市场对百香果果汁需求量逐渐增高,但传统物理压榨法出汁率较低,为提高百香果出汁率,本试验以台农1号百香果为原料,利用超声波辅助复合酶法,通过研究超声温度、超声时间、酶解时间、酶解温度、复合酶配比和加酶量等因素对百香果果汁的影响,在单因素的基础上进行正交试验,以出汁率和感官评价结果为依据,得出最佳加工工艺,并对运用该工艺提取的百香果果汁进行品质分析。结果表明:最佳工艺为超声温度为20℃、超声时间为30 min、酶解时间为60 min、酶解温度为20℃、酶配比为1:1(果胶酶:纤维素酶)、加酶量为0.24%,与直接压榨果肉相比出汁率提高23.29%,维生素C含量损失率降低了20.88%,可溶性糖含量提升了42.41%、总抗氧化能力、ABTS清除能力和DPPH清除能力和总酚含量分别提升了55.23%、7.73%、73.18%和37.61%,对氨基酸含量的影响差异不显著。因此,采用超声波辅助酶法处理百香果可以显著提升百香果出汁率及抗氧化能力。