Study on screening and cultivation conditions of xylanase-producing alkalophilic bacterial

Xylanisthemajorcomponentofplantcellwallsandthemostabundantpentosaninnature.Xylancontainsβ 1,4 linkedd xylosebackboneswitharabinose,4 O methyl d glucuronicacid,andaceticacidsubstituents[1 ] .Thexylan degradingen zymesincludeendoxylanase(1,4 β d xylanxylanohydrolase;EC3.2 .1.8)and β xylosidase (1,4 β d xylanxylanohydrolase;EC 3.2 .1.37) .xylanasedegradesthexylanbackboneintoxylo bioseandxylooligosaccharides,whileβ xylosidasereleasesxylosylresiduesfromthenonreducingendsofxylooligosaccha…     

Role of Cys221 and Asn116 in the zinc-binding sites of the Aeromonas hydrophila metallo-β-lactamase

The CphA metallo--lactamase produced by Aeromonas hydrophila exhibits two zinc-binding sites. Maximum activity is obtained upon binding of one zinc ion, whereas binding of the second zinc ion results in a drastic decrease in the hydrolytic activity. In this study, we analyzed the role of Asn116 and Cys221, two residues of the active site. These residues were replaced by site-directed mutagenesis and the different mutants were characterized. The C221S and C221A mutants were seriously impaired in their ability to bind the first, catalytic zinc ion and were nearly completely inactive, indicating a major role for Cys221 in the binding of the catalytic metal ion. By contrast, the binding of the second zinc ion was only slightly affected, at least for the C221S mutant. Mutation of Asn116 did not lead to a drastic decrease in the hydrolytic activity, indicating that this residue does not play a key role in the catalytic mechanism. However, the substitution of Asn116 by a Cys or His residue resulted in an approximately fivefold increase in the affinity for the second, inhibitory zinc ion. Together, these data suggested that the first zinc ion is located in the binding site involving the Cys221 and that the second zinc ion binds in the binding site involving Asn116 and, presumably, His118 and His196.Received 3 March 2003; received after revision 4 August 2003; accepted 25 August 2003     

Changes in calpastatin localization and expression during calpain activation: a new mechanism for the regulation of intracellular Ca<Superscript>2+</Superscript>-dependent proteolysis

The amount of calpastatin directly available in cytosol is under the control of [Ca²⁺] and [cyclic AMP]. Prolonged calpain activation also promotes degradation of calpastatin. The fluctuation of calpastatin concentration in cell soluble fraction is accompanied by an initial decrease in calpastatin gene expression, followed by a fivefold increase in its expression when the inhibitor protein is degraded. This process can be conceptualized as a mechanism to regulate calpastatin availability in the cell. This conclusion is supported by the fact that calpain, the other component of this proteolytic system, undergoes changes in its levels of expression in a much more limited manner. Furthermore, this process can be observed both in cells exposed to different natural stimuli, or in other cell lines. Modification of calpastatin gene expression might represent a new tool for the in vivo control of the regulatory machinery required for the modulation of Ca²⁺-dependent proteolysis.Received 18 July 2003; received after revision 3 September 2003; accepted 23 September 2003     

紫外线诱变康宁木霉提高酶活力的研究

通过对康宁木霉菌种的诱变、筛选和固体发酵培养,研究了培养基、培养时间、诱变时间及pH值对诱变菌所产酶的活力的影响．结果表明,康宁木霉经诱变5min,在pH为6．0的玉米秸PDA培养基上培养72h,产生的C1酶和Cx酶的酶活力较高,C1酶的酶活力由73u／g提高到176u／g,增加了2．41倍,Cx酶的酶活力由798u／g提高到2069u／g,增加了2．59倍．     

Fluorescence spectra and enzymatic property of hemoglobin as mimetic peroxidase

0　IntroductionHemoglobin(Hb)isthemajorhemeproteinofredbloodcells(RBCs)andisresponsibleforthetransportofoxygentothetissues.ThefunctionofHbdependsupontheabilityoffer rousironinthehemegrouptobindandreleaseoxygen .Despiteitsprincipalroleasanoxygen carrier,theHbmoleculepossessesdifferentenzymaticactivities[1 ] andamethodwasdevelopedforthedeterminationofHbbasedonitsenzymaticactivityfortheox idationofo phenylenediamine (OPDA)withH2 O2 asanoxi dant[2 ] .Hbasamimeticenzymeofperoxidase ,cancataly…     

不同温度对双齿多刺蚁养殖及其保护酶系的影响

在不同温度下长期养殖双齿多刺蚁,观察其存活率和活动状况,测定了不同温度对其SOD,POD,CAT活力的影响,并进行了保护酶活性和存活率的相关分析.实验表明,在较低和较高温度下其存活率明显低于10-40℃测定值.在40-45℃时,酶的活性随着温度升高急速下降;在-5-10℃时,酶的活性随着温度降低而下降.保护酶活性和存活率密切相关.10℃以下的低温和40℃以上的高温对其有不同程度的伤害,其适宜温区为10-40℃,最适温度为20-30℃.     

2,3-Dihydroxybiphenyl dioxygenase gene was first discovered in Arthrobacter sp. strain PJ3

Bacterium strain PJ3,isolated from wastewater and identified as Arthrobacter sp. bacterium based on its 16S rDNA gene,could use carbazole as the sole carbon,nitrogen and energy source. The genomic library of strain PJ3 was constructed and a positive clone JM109(pUCW402) was screened out for the expression of dioxygenase by the ability to form yellow ring-fission product. A 2,3-dihydroxybiphenyl dioxygenase(23DHBD) gene of 933 bp was found in the 3360 bp exogenous fragment of pUCW402 by GenSCAN software and BLAST analysis. The phylogenetic analysis showed that 23DHBD from strain PJ3 formed a deep branch separate from a cluster containing most known 23DHBD in GenBank. Southern hybridization confirmed for the first time that the 23DHBD gene was from the genomic DNA of Arthrobacter sp. PJ3. In order to test the gene function,recombinant bacterium BL21(pETW-8) was constructed to express 23DHBD. The expression level in BL21(pETW-8) was highest compared with the recombinant bacteria JM109(pUCW402) and strain PJ3. We observed that 23DHBD was not absolute specific. The enzyme activity was higher with 2,3-dihydroxybiphenyl as a substrate than with catechol. The substrate specificity assay suggested that 23DHBD was essential for cleavage of bi-cyclic aromatic compounds during the course of aromatic compound biodegradation in Arthrobacter sp. strain PJ3.     

兔毛纺织品防脱毛技术的研究

对兔毛纤维采用生物酶处理，使其形态结构发生改变，从而提高了兔毛纤维的表面摩擦系数，增加了纤维之间的抱舍力，提高了纤维的可纺性，改善了兔毛纺织品的脱毛现象．     

产结晶纤维素酶的嗜热菌鉴定及其酶学性质

从云南腾冲热海温泉采集的样品中,分离出一株产结晶纤维素酶的嗜热菌TC-1.该菌株呈杆状,革兰氏阴性,生理生化特征分析和16S rDNA序列同源性比对表明它属于亚栖热菌属Meiothermus.结晶纤维素、木聚糖和葡萄糖可以诱导该菌产生结晶纤维素酶,其中结晶纤维素诱导的酶活力最高.对其产生的结晶纤维素酶进行了初步纯化与酶学性质研究,该酶的最适反应温度为70℃,最适pH为6.0,在50℃～70℃和pH5.5～6.5之间酶活力相对稳定.     

采用蛋白酶组合漂白技术对羊毛增白的探讨

根据厂家对羊毛织物白度要求，需要开发一种快速，安全，价廉的羊毛漂白生产技术以满足于生产研究，研究了蛋白关正常氧漂工艺中对织物白度的促进作用及采取正确处理方法是获得良好的必要条件。