顺式环氧琥珀酸水解酶产生菌的筛选及产酶条件

从65株诺卡氏菌中,筛选到一株产顺式环氧琥珀酸水解酶(ESH)的菌株,经鉴定为酒石酸诺卡氏菌(Nocardia tartaricans)SW13-57。在14L发酵罐中通过诱导培养,能在细胞内产生ESH酶活力达120u/g。产酶条件研究表明用丙二醇作碳源,硫酸铵作氮源,顺式环氧琥珀酸作诱导剂,初始pH7.0,温度30℃,通过培养24～30h,产酶量最高。该产酶菌株已被用于固定化细胞方法连续生产L(+)酒石酸。     

棒状杆菌固定化细胞生产L(+)-酒石酸

以卡拉胶为载体,固定化棒状杆菌(Corynebacterium sp.)JZ1菌株细胞,再经活化处理,顺式环氧琥珀酸水解酶(ESH)酶活力总回收率在100%以上。摇瓶反应10批,酶活力没有明显降低。1500L酶柱中连续运行90d,酶稳定性很好。固定化后酶反应的最适温度(45℃)和最适pH(90)没有改变,而热稳定性、pH稳定性增强     

产顺式环氧琥珀酸水解酶的博德特氏菌BK-52的筛选、鉴定及其产酶条件优化

[目的]产顺式环氧琥珀酸水解酶(CESH)新型菌株的筛选、鉴定及其产酶条件优化.[方法]通过电镜、Biolog GN、(G C)含量和16S rDNA序列研究,对筛选菌株进行分类鉴定.通过红外光谱、核磁共振氢谱和碳谱、质谱以及旋光度实验,鉴定纯化产物的结构并优化CESH产酶条件.[结果]本文筛选出一株产CESH的新型菌株,该菌株可将顺式琥珀酸盐转化为D(-)-酒石酸,属于博德特氏菌属,并将其命名为博德特氏菌BK-52.最佳发酵条件为:最佳温度30℃,最佳pH7.0,最佳发酵时间36 h,最佳碳源蔗糖,最佳无机氮源硫酸铵,最佳酶诱导剂顺式环氧琥珀酸二钠.在此最佳条件下,CESH酶活最高达764 U/g生物量.[结论]本文筛选的新菌株博德特氏菌BK-52为D(一)一酒石酸的生产提供了一种新的方法.     

青霉PT95菌株固态发酵产生类胡萝卜素的研究

本文对青霉Penicillium sp. PT95菌株在固态发酵条件下菌核内产生类胡萝卜素进行了初步研究。结果表明,在3种固态发酵培养基中,玉米粉培养基(SMA)比麸皮培养基和棉籽壳培养基更适合于PT95菌株固态发酵产生类胡萝卜素。为了增加菌核干重和提高类胡萝卜素产率,SMA中需要添加氮源、碳源和植物油。在所试的各种氮、碳源中,以硝酸钠和麦芽糖效果最佳。通过正交试验确定了在培养基盐溶液中添加硝酸钠3g/L,麦芽糖10g/L,豆油2.5g/L能使菌核干重由536g/100g提高到970g/100g(干料);类胡萝卜素产率由2149μg/100g提高到5260μg/100g(干料);β-胡萝卜素在类胡萝卜素中的含量由614%提高到71.3%。     

假单胞菌产脂肪酶条件的初步探索

对假单胞菌（Pseudomonassp）2106菌株产脂肪酶条件的初步探索表明,该菌株脂肪酶为组成型,不受油脂类底物的诱导。碳源的种类（单糖、双糖、多糖）和浓度对产酶影响不大,氮源以豆饼粉和玉米浆混合添加最好。最适发酵温度为32℃,摇瓶转速140r／min。实验中还通过正交试验优化了产酶培养基组成。     

微量元素铬载体酵母摇瓶发酵研究

实验选择了啤酒酵母 2 0 16菌株为铬载体酵母生产菌株 ,通过摇瓶发酵实验 ,对三氯化铬的添加工艺进行了研究探索。确定了铬酵母发酵培养的适宜加铬条件 :在发酵开始的8h内分批流加三氯化铬 ,总加入量为 5× 10⁴mol/L ,经过 12h摇瓶发酵可得铬酵母的生物量为 1 5 5 g/ 10 0mL ,酵母含铬量为 1,10 0 μg/ g以上 ,酵母细胞对铬离子利用效率是 6 0 %以上。     

嗜线虫致病杆菌HB310培养基筛选和培养条件优化

嗜线虫致病杆菌Xenorhabdus nematophila是与小卷蛾斯氏线虫Steinernema carpocapsae互惠共生的一种革兰氏阴性菌,对多种农业害虫都具有很高的杀虫活性。本研究对多种影响嗜线虫致病杆菌HB310菌株发酵培养的因素进行了筛选优化。通过单因素试验确定最佳碳源、氮源和无机盐分别为葡萄糖、牛肉蛋白胨和KH2PO4;通过正交实验确定培养基的最优组合为：葡萄糖2 %、牛肉蛋白胨1 %、牛肉浸膏0.3% 和K2HPO4.1 %;最佳摇瓶培养条件为：接种量6%、培养基pH 7.5、摇床转速200 rpm、培养温度28℃和培养时间48 h,在此条件下,嗜线虫致病杆菌HB310菌株生长速率最快,菌液的杀虫活性最高。     

一株产木聚糖酶链霉菌的鉴定及发酵产酶*

以木聚糖为唯一碳源,筛选出一株高产木聚糖酶生产菌株。该菌株经形态特征、 培养特征、生理生化和细胞壁组分分析等实验,鉴定为卷须链霉菌（Streptomyces cirratus).摇瓶发酵产酶实验表明：培养基最佳初始pH值为6．0;玉米芯水不溶木聚糖和蛋白胨分别是最佳的碳源和氮源;添加0．5％吐温80使得木聚糖酶活力提高到原来的2．5倍,发酵液最高酶活达到623u／mL。     

富锌酵母的选育及培养条件研究

对402株不同种属的酵母菌株进行了出筛、复筛、单倍体分离、诱变,从亲株Y-6-16-18（a met）accharomyces cerevisiae）和Y378-4-15（α-leu）（Sacharomyces kluyveri）的杂交菌株中选育到一株富锌酵母菌株（编号为ZGH374）。并初步优化了发酵条件：培养基为80g/L糖浓度的麦芽汁、10g/L蛋白胨、锌添加量400μg/mL,pH 6.0,装液量40mL/250mL三角瓶,接种量10%（V/V）,培养起始添加锌盐,培养时间30h。在优化的条件下,杂交菌株ZGH374的生物量（细胞干重）达到14.3g/L,细胞锌含量可达到9.3mg/g,锌总含量达到了133mg/L。     

十三碳二元羧酸发酵技术的研究

以一株热带假丝酵母菌(Candida tropicalis) SP1为出发菌株,经紫外线反复诱变获取一株难以同化烷烃的突变株SPUV56,摇瓶培养5d平均产酸量达72g/L,较出发菌株提高了1.25倍,并利用突变株SPUV56在137L自控罐上扩试,补加醋酸盐发酵,144h产酸量达153g/L,比不加醋酸盐发酵提高了29.7%。采用提 高搅拌混合效果和低溶解氧发酵过程控制方法,可有效地提高菌体的产酸能力,在20m3发酵罐中发酵生产十三碳二元羧酸,总培养时间144h,产酸量可达172g/L,放罐体积15.0m3,产量为2.25t。     

环氧化物水解酶产生菌的筛选及产酶条件研究

手性环氧化物至少含有一个手性碳,通过选择性开环和官能团转换,可以方便地合成许多有价值的手性化合物,在制药、农药、香料、精细化学品工业上有着极其重要的应用价值[1]。因此,手性环氧化物的合成一直是一个重要的研究课题。     

一株乙腈降解菌的鉴定及其生物学特性

【目的】明确乙腈降解菌BX2的分类地位及生物学特性,评价其处理含乙腈废水的可行性。【方法】通过形态特征、生理生化特性以及系统发育分析对菌株BX2进行鉴定。考察温度、初始pH及接种量等因素对菌株生长的影响,确定菌株的最佳生长条件及在该条件下的乙腈降解能力。测定菌株BX2对NaCl的耐受能力。【结果】乙腈降解菌BX2的形态特征及生理生化特性与紫红红球菌(Rhodococcus rhodochrous)最相近。其16S rRNA、gyrB、secA1基因序列与紫红红球菌的相似性分别为99.37%、99.29%、97.87%。最佳生长条件为35℃,初始pH 7.5,接种量1%。此条件下,菌株BX2在16 h内对浓度为800 mg/L乙腈的降解率为95.87%。菌株BX2在NaCl含量高于6%的培养基中无法生长。【结论】菌株BX2被鉴定为紫红红球菌。该菌株有较强的环境适应能力,可降解高浓度乙腈,在含氰废水的生物修复中有很好的应用前景。     

Expression and inducibility of endosulfan metabolizing gene in Rhodococcus strain isolated from earthworm gut microflora for its application in bioremediation

The metabolizing potential of a bacterial strain Rhodococcus MTCC 6716, isolated from the gut of an Indian earthworm (Metaphire posthuma) was studied for endosulfan bioremediation. In the present work, the optimum conditions for the maximum growth, kinetic of endosulfan degradation, regression equation, half life and correlation coefficient were studied. Endosulfan induced alterations in the expression of mRNA and protein of specific endosulfan metabolizing marker gene (Esd) was studied. Maximum growth of bacteria was observed at pH 7.0, 30°C and 0.085 M sodium chloride concentration in a liquid culture medium. Endosulfan was degraded by Rhodococcus strain up to 97.23% within 15 days without producing toxic metabolite and with strong correlation coefficient (-0.728) and half life 5.99 days. Endosulfan degradation was mediated through gene(s) present in genomic DNA. Expression of marker gene was found endosulfan concentration dependent. The results suggest that this novel strain (Rhodococcus) may be utilized for bioremediation of endosulfan.     

降解鸡毛真菌板蜡蚧的鉴定及产酶培养优化

鉴定降解鸡毛真菌并通过单因素和正交实验优化其产角蛋白酶发酵培养条件.从加入鸡毛粉钓饵的医院花坛土中分离筛选获得3株角蛋白高效降解真菌,利用形态学和分子系统学鉴定均为板蜡蚧(Lecani-cillium testudineum).单因素实验表明,对优选菌株1Y2-12产酶能力具促进作用的碳源为乳糖,氮源为酵母膏,无机离子...     

Cloning and expression of the s-triazine hydrolase gene (trzA) from Rhodococcus corallinus and development of Rhodococcus recombinant strains capable of dealkylating and dechlorinating the herbicide atrazine.

We used degenerate oligodeoxyribonucleotides derived from the N-terminal sequence of the s-triazine hydrolase from Rhodococcus corallinus NRRL B-15444R in an amplification reaction to isolate a DNA segment containing a 57-bp fragment from the trzA gene. By using the nucleotide sequence of this fragment, a nondegenerate oligodeoxyribonucleotide was synthesized and used to screen a genomic library of R. corallinus DNA for fragments containing trzA. A 5.3-kb PstI fragment containing trzA was cloned, and the nucleotide sequence of a 2,450-bp region containing trzA was determined. No trzA expression was detected in Escherichia coli or several other gram-negative bacteria. The trzA gene was subcloned into a Rhodococcus-E. coli shuttle vector, pBS305, and transformed into several Rhodococcus strains. Expression of trzA was demonstrated in all Rhodococcus transformants. Rhodococcus sp. strain TE1, which possesses the catabolic gene (atrA) for the N-dealkylation of the herbicides atrazine and simazine, was able to dechlorinate the dealkylated metabolites of atrazine and simazine when carrying the trzA gene on a plasmid. A plasmid carrying both atrA and trzA was constructed and transformed into three atrA- and trzA-deficient Rhodococcus strains. Both genes were expressed in the transformants. The s-triazine hydrolase activity of the recombinant strains carrying the trzA plasmid were compared with that of the R. corallinus strain from which it was derived.     

一株产D-海因酶菌株的鉴定及离子束诱变选育

首次报道了产D-海因酶的巨大芽孢杆菌,通过对该菌进行离子束诱变,获得酶活最高增加3倍的突变株M5,对M5的产酶条件进行优化,得到最佳的发酵条件为玉米浆1.5%,葡萄糖1%,油酸1.5%,氯化钠0.5%,并添加50 mg.L-1的Mn2+、Zn2+及500 mg.L-1的Mg2+,pH8.0,30℃发酵24 h,酶活力可达到每毫升2.119 U,比优化前突变株提高了300%,比出发菌株提高了850%。     

A novel enantioselective epoxide hydrolase for (R)-phenyl glycidyl ether to generate (R)-3-phenoxy-1,2-propanediol

Bacillus sp. Z018, a novel strain producing epoxide hydrolase, was isolated from soil. The epoxide hydrolase catalyzed the stereospecific hydrolysis of (R)-phenyl glycidyl ether to generate (R)-3-phenoxy-1,2-propanediol. Epoxide hydrolase from Bacillus sp. Z018 was inducible, and (R)-phenyl glycidyl ether was able to act as an inducer. The fermentation conditions for epoxide hydrolase were 35°C, pH 7.5 with glucose and NH₄Cl as the best carbon and nitrogen source, respectively. Under optimized conditions, the biotransformation yield of 45.8% and the enantiomeric excess of 96.3% were obtained for the product (R)-3-phenoxy-1,2-propanediol.     

Isolation and identification of a novel Rhodococcus sp. ML-0004 producing epoxide hydrolase and optimization of enzyme production

Rhodococcus sp. ML-0004, a novel strain for producing epoxide hydrolase, was isolated from soil in this study. The epoxide hydrolase can catalyze the stereo-specific hydrolysis of cis-epoxysuccinic acid to generate l(+)-tartaric acid. By examining physiological, biochemical characteristics and comparing its 16S rDNA gene sequence, it was identified as Rhodococcus opacus, and named R. opacus ML-0004. The optimal conditions for epoxide hydrolase production from R. opacus ML-0004 were also investigated. Propanediol and (NH₄)₂SO₄ were selected as carbon source and nitrogen source, respectively, for the production of R. opacus ML-0004 epoxide hydrolase. The optimal conditions for epoxide hydrolase production were fermentation temperature = 28 °C, pH 7.0, and cultivation time = 26 h. Under these conditions, the maximum epoxide hydrolase activity reached 10.5 U mL⁻¹.     

产D-阿拉伯醇菌株的筛选、鉴定及其产D-阿拉伯醇条件的优化

摘要：【目的】产D-阿拉伯醇的耐高渗酵母的筛选、鉴定和产D-阿拉伯醇条件的优化。【方法】通过电镜、Biolog GN、(G+C)含量和26S rDNA D1/D2区序列分析法对所获得的菌株进行了描述。通过红外光谱、核磁共振氢谱和碳谱、质谱以及旋光度实验鉴定纯化产物的结构。通过单因素实验优化产D-阿拉伯醇的发酵条件。【结果】本文筛选得到一株产D-阿拉伯醇的新型菌株,经鉴定属于假丝酵母属并命名为Candida sp. H2。200 mL摇瓶发酵生产D-阿拉伯醇的单因素优化实验表明,最适发酵条件为：葡萄糖250