产几丁质酶菌株SWCH-6的筛选、鉴定及其产酶条件的优化研究

从汕头海湾养殖区域的海底沉积物中分离到1株几丁质酶活性较高的菌株, 命名为SWCH-6, 根据菌株的形态特征、生理生化特征和16S rDNA 序列, 确定该菌株为嗜水气单胞菌(Aeromonas hydrophlilla)。采用单因素优化方法结合正交实验, 得到菌株SWCH-6产几丁质酶的最佳发酵条件：胶体几丁质25.0 g/L, 胰蛋白胨10.0 g/L, 陈海水1.0 L, pH 8.5, 32℃, 150 r/min培养72 h; 在该条件下酶活力达0.39 U/mL。此外, 菌株所产几丁质酶的最适催化pH 5.0; 最适催化温度为40℃; Cu2+、Fe3+及表面活性剂Tween-80能增强该酶的催化活性; Zn2+、Mn2+及表面活性剂SDS、洗衣粉对该酶的催化活性有抑制作用, 与其它几丁质酶存在着一些不同。     

产几丁质酶的苏云金杆菌菌株筛选及酶合成条件研究

从本室保存的 64株苏云金芽孢杆菌中 ,筛选出一株几丁质酶活力较高的菌株WB 50。产酶条件研究表明 :在pH 7.0的基础培养基中添加 2 .0 %的细粉几丁质 ,1.0 %的酵母膏 ,2 2 0rpm 30℃下培养 72小时 ,几丁质酶的产出最大。     

产几丁质酶菌株SWCH-6的筛选、鉴定及其产酶条件的优化研究

从汕头海湾养殖区域的海底沉积物中分离到1株几丁质酶活性较高的菌株,命名为SWCH-6,根据菌株的形态特征、生理生化特征和16S Rdna序列,确定该菌株为嗜水气单胞茵(Aeromonas hydrophlilla).采用单因素优化方法结合正交实验,得到菌株SWCH-6产几丁质酶的最佳发酵条件:胶体几丁质25.0g/L,胰蛋白胨10.0g/L,陈海水1.0L, Ph 8.5,32℃,150 r/min培养72h;在该条件下酶活力达0.39U/Ml.此外,菌株所产几丁质酶的最适催化Ph 5.0;最适催化温度为40℃;Cu2、Fe3及表面活性剂Tween-80能增强该酶的催化活性;Zn2 、Mn2 及表面活性剂SDS、洗衣粉对该酶的催化活性有抑制作用,与其它几丁质酶存在着一些不同.     

几丁质酶产生菌的筛选及产酶条件的研究*

从301株几丁质降解菌包括细菌、放线菌和真菌中,筛选出一株产几丁质酶活力较高的链霉菌A048(Streptomycessp．A048)。其产酶的适宜条件是,培养温度30℃,培养基pH7．0～7．5,碳源几丁质,氮源NH4NO3,体积溶氧系数kd值为1．56×10-6mol／(mL·min·atm),振荡培养120h,产酶可达15．1U／mL。     

芽胞杆菌菌株产几丁质酶发酵条件的研究

从采自大连地区24份土样中分离筛选到一株产几丁质酶活性较高的芽胞杆菌(Bacillus sp.)B-41,该菌株产几丁质酶最适的发酵条件为：碳源为胶体几丁质,氮源为酵母浸汁,pH7．2,温度42℃,振荡培养4天。     

海洋低温几丁质酶菌株筛选、鉴定及酶谱分析

以胶体几丁质为唯一碳源,从大连渤海湾的底泥样品中分离到1株高产低温几丁质酶的海洋细菌,命名为DL-06。由菌株的形态特征结合16S rDNA系统发育分析,初步确定该菌株属于交替假单胞菌(Pseudoalteromonas sp. DL-06)。该菌株经30 h摇瓶发酵后测定粗酶液几丁质酶酶活为9.184 U/mL,最适反应温度为15 ℃,60 ℃孵育1 h仍保持50%以上的酶活性,表明该低温酶具有一定热稳定性。经SDS-PAGE及酶谱分析,该菌株能够产生至少3种以上不同分子质量的几丁质酶组分。Pseudoalteromonas sp. DL-06产几丁质酶在低温下高活性与热稳定特点,使其具有潜在的工业应用价值。     

几丁质酶产生菌筛选鉴定及产酶性能研究

从土壤样品中筛选得到一株高产几丁质酶菌株C65-2,经形态学观察和18S rDNA序列测定,鉴定为Aspergillus fumigatus,对产酶培养基进行初步优化,测得最高酶活可达6.9U/ml,酶活力较优化之前提高了210%。酶学性质研究表明该几丁质酶分子量约为20kDa,酶在60℃下保温50min酶活降为0,最适酶反应温度是55℃,酶反应最适pH为7.0,Mg2+,Cu2+对酶反应有促进作用,Fe3+对酶反应有抑制作用。     

Serratia sp. PS-2产几丁质酶发酵条件研究

采用平板透明圈法,从海洋红树林根泥筛选得到一株产几丁质酶较强沙雷氏菌株Serratia sp. PS-2.为实现工业化生产几丁质酶,考察了以几丁质为诱导剂,碳源、氮源、NaCl 浓度、培养温度、时间、培养液酸碱度、通气量和搅拌速度等对产酶的影响,得到了优化的发酵条件.5L发酵罐实验表明,该菌株可在发酵72h内达到产酶高峰,最高酶活力可达到0.68 U/mL.     

一株产漆酶菌株的筛选鉴定和发酵条件的研究

以愈创木酚为底物,采用平板筛选法筛选得到一株产漆酶菌株WS1-2,形态学特性表明该茵属于绿色木霉.对产酶条件的初步研究结果表明,WS1-2菌株的产酶高峰期出现在接种培养后的第4d.与蔗糖、乳糖、半乳糖和可溶性淀粉相比,以葡萄糖为碳源时,发酵上清液的漆酶活力明显要高,最大值达230U/L.以NH4Cl为氮源,最有利于WS1-2漆酶的产生,漆酶活力最高可达到234U/L.0.01mmol/L的愈创木酚和ABTS对WS1-2产漆酶有明显的诱导作用,3～5mg/L的Tween-80可以明显提高WS1-2的产酶水平.     

产果胶酶菌株的筛选

从不同来源的生物样品中分离到产果胶酶菌株,采用刚果红染色法和十六烷基三甲基溴化铵沉淀法进行初筛,通过摇瓶发酵试验进行复筛,最终获得2株果胶酶活力较高的细菌G16和G25,其酶活力分别达到6．37万U／mL和6．84万U／mL。     

N—乙酰氨基己糖苷酶产生菌的筛选与产酶条件

About 1200 strains of microorganisms were screened including fungi, actinomyces, and bacteria, in which 237 strains producing the enzyme desired. The results showed that the beta-GlcNAcase and beta-GalNAcase always co-existed in one strain, though may be in different ratio. From strains mentioned above the authors screened out a potent beta-N-acetylhexosaminidase producing strain, Aspergillus tamarii S215, from the soil sample. The optimal conditions for enzyme production were as follows: the microorganisms was inoculated in a 5% wheat bran suspension, cultured at 28-30 degrees C on shaker for 5-6 days. The productivity can be moderately enhanced by the addition of cellobiose or glucosamine or galactosamine or by the extra supplement of (NH4)2SO4 and NH4NO3 as N sources. In the culture filtrate of Asp. tamarii, the alpha, (beta)-galactosidase, beta-glucosidase, alpha-mannosidase and beta-fucosidase were also found.     

产几丁质酶褐色喜热裂孢菌的分离鉴定及产酶特性初步研究

采用选择性培养基从土壤中分离到1株产几丁质酶的微生物菌株YX,经形态和分子鉴定为褐色喜热裂孢菌(Thermobifida fusca)。进一步在摇瓶中比较了T.fusca YX在纤维二糖、几丁质、或羧甲基纤维素钠为碳源的培养基中的产酶特性,YX菌株在5 L发酵罐中以几丁质为碳源的培养基发酵到22 h左右时发酵液几丁质酶活即可达到1.7 U/m L。本文首次报道褐色喜热裂孢菌能够产生几丁质酶,具有潜在的应用价值。     

Purification,cDNA cloning,and characterization of LysM-containing plant chitinase from horsetail (Equisetum arvense)

Chitinase-A (EaChiA), molecular mass 36 kDa, was purified from the vegetative stems of a horsetail (Equisetum arvense) using a series of column chromatography. The N-terminal amino acid sequence of EaChiA was similar to the lysin motif (LysM). A cDNA encoding EaChiA was cloned by rapid amplification of cDNA ends and polymerase chain reaction. It consisted of 1320 nucleotides and encoded an open reading frame of 361 amino acid residues. The deduced amino acid sequence indicated that EaChiA is composed of a N-terminal LysM domain and a C-terminal plant class IIIb chitinase catalytic domain, belonging to the glycoside hydrolase family 18, linked by proline-rich regions. EaChiA has strong chitin-binding activity, however, no antifungal activity. This is the first report of a chitinase from Equisetopsida, a class of fern plants, and the second report of a LysM-containing chitinase from a plant.     

褐藻降解菌群筛选及其复合酶固定化研究

以大连地区的褐藻为材料,筛选了褐藻降解菌群,其在30℃、pH7．5条件下培养74h时,酶活达1．883IU／mL。利用硅藻土吸附法经冷冻干燥制备了固定化复合酶,分析了复合酶系的酶学性质,其最适反应温度为45℃,并在40℃～55℃范围内具有良好的热稳定性;最适pH为7．5,并在pH7．0～8．5之间pH稳定性良好。利用固定化酶进行褐藻酸钠制备,提取率达48．3％,粘度为2．9Pa·s,与传统方法相比均有显著提高,为工程化生产褐藻酸钠提供一定基础。     

产海因酶的菌种筛选和产酶条件的研究

利用5-苄海因作为唯一氮源法筛选高产海因酶的菌种,从本实验室保存的221株菌种中筛选出12株具有不对称水解5-苄海因生成N-氨甲酰基-苯丙氨酸的菌株,其中假单胞菌（Pseudomonassp.)X4-49具有较高的产酶活力,对此菌的产酶条件的研究表明,产酶的最佳碳源为甘油,最佳氮源为蛋白胨,最佳诱导物为苄海因,尿嘧啶,苄海因作为诱导物的有效浓度为0.2%,产酶的最适培养基的初始pH为7.0。培养条件为33℃,13h。     

昆虫几丁质酶及其在害虫防治中的应用

几丁质是昆虫重要的结构性组分,在昆虫生长发育的各个时期都需要一定量的几丁质来维持其代谢平衡.昆虫几丁质酶可以降解昆虫体壁和围食膜中的几丁质,作为一种潜在的生物杀虫剂在害虫防治方面具有广阔的应用前景.随着对昆虫几丁质酶研究的不断深入,目前已克隆到了30余种昆虫几丁质酶,并应用于转基因作物和基因工程微生物中,对害虫具有一定...     

产柚苷酶菌株B04的分离及产酶特性研究

柚苷酶作为金柚和柑桔果实中主要苦味物质的分解酶有望在深加工生产中得到应用。本研究在金柚树林下土壤和金柚皮腐烂液中分离得到产柚苷酶菌株。研究证明采用添加低浓度蔗糖作为碳源的高层试管法是可行的初筛方法;复筛得到菌株B04,产酶需要诱导,酶底物柚苷和产物鼠李糖均有诱导作用,其中柚苷诱导作用更好,而且添加量为0.02%时菌株产酶能力较强;B04发酵类型为非生长偶联型,30℃,175r/min旋转摇床培养,60h菌体量最大,96h酶活力最高,达到342U/mL。     

Effect of environmental factors on expression and activity of chitinase genes of vibrios with special reference to Vibrio cholerae

AIMS: The aim of this study was to investigate the distribution and inducibility of chitinase genes in vibrios and the effect of environmental factors on the expression level and activity of chitinase genes in Vibrio cholerae strains. METHODS AND RESULTS: Chitin agar plate assays showed that V. cholerae strains were more chitinolytic than non-cholerae vibrios. All of the identified or putative chitinase genes were expressed in V. cholerae (four strains) but not in non-cholerae vibrios (seven species/strains) under standard laboratory growth conditions. In non-cholerae vibrios, these genes were induced by chitin, its monomer N-acetyl-d-glucosamine and on exposure to rabbit intestine, while in V. cholerae strains, these genes showed significant variation in expression levels. To study the effects of environmental factors on the expression and activity of chitinase genes in V. cholerae, bacteria were cultured in different pH, temperature, sodium chloride and nutrients. RT-PCR analysis showed that lower temperatures and higher pH, salinity and nutrition favoured expression of these genes, while their activity increased under higher nutrition content and salinity. CONCLUSIONS: Chitinase genes are distributed in all the relatively small number of strains studied here, and biotic and abiotic factors have significant role in the induction, expression level and activity of this gene family in vibrios. SIGNIFICANCE AND IMPACT OF THE STUDY: Chitinases have important applications especially in recycling of chitin. Vibrios can be used as chitinolytic agents, using suitable culture conditions that maximize the expression and activity of these genes.     

昆虫几丁质酶及其在植物保护中的应用

本文从生理学、生物化学和分子生物学３个方面综述了昆虫几丁质酶的研究进展,并概述了它在害虫防治、主要是在抗虫育种中作为重要的基因源中的应用。e