产几丁质酶菌株的分离与鉴定及产酶条件优化

从土壤中分离到一株产几丁质酶的细菌LL-C.该菌经16S rDNA序列同源性分析及形态培养特征、生理生化特征分析,鉴定为Luteibacter rhizovicinus,属于黄单胞菌科(Xanthomonadaceae).以不同发酵时间、碳源、氮源、起始pH值、培养基装液量和培养温度为因素,考察LL-C菌株产几丁质酶的优化条件.结果表明,最有利于该菌产几丁质酶的条件:碳源为胶体几丁质、氮源为(NH4)2SO4,培养基起始pH值为4.5,培养基装液量为30 mL,培养温度为32 ℃,振荡培养时间为7 d,此优化条件下,摇瓶产酶活力为115.02 μkat·L-1.     

海洋弧菌Z010产生几丁质酶的发酵条件研究

以海洋弧菌Ｚ０１０菌株为对象,研究了其产生几丁质酶的发酵条件。研究表明,其产酶的最佳发酵条件为：２２１６Ｅ液体培养基中含０．５％（ｗ／ｖ）的胶体几丁质,０．５％（ｗ／ｖ）的蛋白胨,培养起始ｐＨ值为７．０,２５℃摇床２００ｒ／ｍｉｎ培养３６ｈ,几丁质酶产出最大。     

纤溶酶高产菌株筛选及其液体发酵条件研究

研究实验室自主分离的2株纤溶酶高产菌株发酵条件,采用4因素、3水平的正交试验设计方法对液体发酵培养基的碳源、氮源、无机盐进行了优化,并对培养基初始pH、发酵时间、发酵温度对产酶量的影响进行了测定.结果表明:菌株1液体发酵合适的产酶培养基配方为大豆粉的质量分数为6%,葡萄糖2%,CaCl2 0.06%,MgSO4 0.07%;菌株2液体发酵合适的产酶培养基配方为大豆粉的质量分数为6%,玉米淀粉2%,CaCl2 0.06%,MgSO4 0.07%.培养基初始pH均为6.5时酶活性最高;发酵温度均以38℃最好;而菌株1和菌株2的适宜发酵时间分别为48～84 h和60～84 h.在优化条件下,菌株1、菌株2液体发酵最高酶活分别为306.46 IU/mL和319.76 IU/mL。     

南海海域几丁质降解菌的筛选及其特性研究

从南海海域水样和沉积物中分离并筛选到两株几丁质降解菌,分别为SCSS04和SCSWE13.16S rDNA序列分析结果表明,菌株SCSS04属于芽孢杆菌属,菌株SCSWE13属于弧菌属.两株菌几丁质酶的产生均需要几丁质的诱导,并在富营养培养基中产酶量较高.菌株SCSS04在培养第9天产酶量达到最高,所产生的几丁质酶最适反应温度为40～50℃.菌株SCSWE13属于低温酶,在培养第7天产酶量达到最高,所产生的几丁质酶最适反应温度为20～28℃.     

土壤产几丁质酶菌株的筛选鉴定及产酶条件

利用几丁质为碳源,从土壤中筛选出3株产几丁质酶菌株,其中酶活最高的为一株革兰氏阴性菌.对该菌株应用16S rDNA法进行鉴定,结果为嗜麦芽窄食单胞菌(Stenotrophomonas maltophilia).通过单因素优化法和均匀设计法实验,结果表明,以质量分数0.5%的胶体几丁质为碳源,质量分数1.0%的蛋白胨为氮源,及30℃和pH值为7.2,发酵60 h的条件是菌株的最合适产酶条件.此时,胞外几丁质酶酶活力最高.     

纳豆激酶产生菌的固体发酵参数优化

对影响纳豆激酶产生菌固体发酵时产酶影响因子如碳源/氮源、含水量、温度、pH和培养时间等进行了优化.实验结果表明,菌株1适宜的固体发酵产酶培养基豆粕:麸皮比为3∶1;菌株2适宜的固体发酵产酶培养基豆粕:麸皮比为1∶1时产酶活性最高;菌株1适宜的培养基含水以50%最好,菌株2以70%最好;培养基初始pH均在7.0时酶活最高;发酵温度均以25℃最好,不易超过30℃;两个菌株的适宜发酵时间分别为36 h(菌株1)和72 h(菌株2).在优化发酵条件下,两个菌株单位发酵物中纤溶酶平均酶活力可分别达到1 407.25 U/g(菌株1)和953 U/g(菌株2).     

地衣芽孢杆菌高α-乙酰乳酸脱羧酶活力菌株的筛选

以从土壤中分离出的9株地衣芽孢杆菌为源菌株,通过α-乙酰乳酸脱羧酶活力实验发现BL-4的产酶活性最好,通过UV和NTG对BL4进行复合诱变,获得了一株产酶活性比出发菌株BL-4高1.22倍的α-乙酰乳酸脱羧酶高产菌株,并研究了该菌株的最佳发酵产酶条件是培养温度30℃、发酵时间36 h、发酵液起始pH 6.5。     

酸性环境中木聚糖酶高产菌株的筛选及鉴定

从pH 5.0的酸性土壤中筛选出一株木聚糖酶高产菌株A4,菌体固态发酵产酶条件优化表明,最佳发酵培养基配方为：麸皮37.79％,玉米芯9.10％,NH4NO3 0.51％,MnSO4 1.60％,水50％,接种量2.0％,最适发酵温度28～32 ℃,发酵培养48～52 h,木聚糖酶活力最高达到750 U/g（碳源）.该菌株所产木聚糖酶的最适pH为4.8,比野生黑曲霉的pH值低.通过生长形态和分子生物学方法相结合的鉴定该菌株为黄曲霉.     

植物乳杆菌的产酸分析

对植物乳杆菌的产酸性能进行了初步研究, 研究结果显示: 该菌株的产酸速度快, 转入培养基后pH即快速下降, 在32h时pH趋于稳定; 属于同型乳酸发酵; 在起始pH为6.0时发酵及产酸能力强.     

海洋链霉菌产生几丁质酶的研究

从闽南地区潮间带红树林根际海泥中分离到能产生几丁质酶(Chitinase)的放线菌(Actinomyces)300余株,研究了各种诱导物、碳氮源及培养条件对链霉菌S-128(Streptomyces sp.S-128)菌株几丁质酶生物合成的影响,结果表明,该菌株几丁质酶的合成能被多种几丁质所诱导,是一种诱导酶,添加氮源尤其是有机氢源能促进酶的合成,而碳源却没有这种作用,基本培养基中加入2.O％脱矿质几丁质和1.5％玉米浆,在28℃发酵5d,发酵液的几丁质酶活性可达290u/ml,该酶对几丁质的水解反应显出较宽的pH范围(pH3～8)和较好的pH稳定性,最适pH5.0,在一定的温度范围内,该酶的活性随温度的上升而增高,并以65℃的酶活性最高。     

CHARACTERISTICS OF QUATERNARY AMMONIUM RESISTANT BACTERIA FROM A FINE PAPERMACHINE SYSTEM

To evaluate the biocide effect of quaternary ammonium chloride (QAC), a whitewater sample was taken from a fine papermachine headbox. By plate spreading method, 51 strains of facultative anaerobe were isolated morphologically. Then the strains were separately transferred to basal medium and were incubated before the beginning of log phase. To identify strains with different QAC resistance, 30-120 ppm of N-Alkyl-benzyl-dimethyl ammonium chloride were added to basal medium.Biocide effect was investigated by comparison of bacterial growth, which can be monitored by 600 nm light absorbance of basal medium suspension. Among 51 strains, only 2 strains can Survive for QAC concentration up to 120 ppm. API20E aud 16S rRNA gene sequencing technique were applied to identify two strains with highest (120 ppm)QAC resistance. One strain (HB22)was identified as Morganella morganii. HB22 can resist QAC concentration up to 150 ppm. HB22 is Gram-negative rod, motile with flagella, catalase positive, oxidase negative, Indole positive, H2S production negative, facultative anaerobe. HB22 has optimum growth condition of 35℃ and pH 7.0.HB22 can catabolize only glucose and D (+)-Mannose.The other (HB45) was identified with high similarity among Pseudomonas cf. monteili or Pseudomonas mosselii or Pseudomonas putida. HB45 can resist QAC concentration up to 200 ppm.HB45 is Gram-negative rod, motile with flagella,catalase positive, oxidase negative, Indole negative,H2S production negative, aerobe. HB45 has optimum growth condition of 30℃ and pH 7.4. HB45 can catabolize L-Arabinose, L(+)-Rhanmose, D (+)-Mannose, Glucose, Glycerol, Lactose, Maltose,Raffmose, Xylose, Cellulose and Xylan. The implication of this work to paper industry is also discussed.     

处理厨余垃圾的高温菌剂研制及其降解性质研究

利用高温菌耐热嗜热的特性和有机物好氧分解的基本原理,从厨余垃圾处理系统中分离筛选到6株在65 ℃能产生淀粉酶、脂肪酶、蛋白质酶及纤维素酶的高温高效菌种.经鉴定,所分离得到的高温菌株都有芽孢,属于兼性细菌.经安全性检测6株高温菌中未检测到沙门氏菌和志贺氏菌等致病菌,说明分离到的高温菌不存在致病性因素.通过最佳菌株组合后对厨余垃圾进行降解试验,最终确定4株（HB1,HB2,HB4和HB6）制成高温菌剂.将该菌剂在100 kg厨余垃圾处理机上进行降解试验,结果表明：一次性投入5%菌剂后升温至65℃,24 h内对厨余垃圾中粗脂肪和粗纤维有明显的降解效果,降解效率分别为30.7%和11.3%,粗蛋白含量增加了9.5%.继续对厨余垃圾处理48 h,其有机物分解率几乎没有得到提高,说明大部分有机物在24 h内即可被分离出的高温菌剂所降解.     

高产聚-3-羟基丁酸重组大肠杆菌的筛选

将能以葡萄糖为唯一碳源积累P(3HB)的工程菌进行优化筛选,获得6个较高产的菌株,PHB含量最高为69.1%.经丙酸驯化,虽然没有PHBV的积累,但PHB的量均有增加,最高可达71.13%.另外,抗生素对菌体生长及PHB积累有重要影响.     

Cloning and Characterization of UL34 Gene of Pseudorabies Virus HB Isolate

In this study, China isolate HB of pseudorabies virus(PRV) was confirmed and genotypically characterized by amplifying and sequencing of partial UL34, a conservative gene involved in the egress of nucleocapsids from the nucleus, for phylogenetic analysis. The open reading frame(orf) of UL34 of PRV HB isolate is composed of 786 nucleotides, which encoded 262 amino acids. In addition, a potential transmembrane domain(241-260 aa) and 11 potential phosphorylation sites were also found in the UL34 of PRV HB isolate. Multiple amino acids alignment indicated that UL34 proteins of PRV strains derived from different geographic origins were highly conservative, but some mutations were also found. Phylogenetic analysis based on UL34 protein indicated that PRV HB strain was evolutionarily distinct from other recent China strains sequenced so far, forming a single clade within the phylogeny. Moreover, PRV HB isolate had close evolutionary relationship with Bo HV-1 and Bo HV-5 within the Alphaherpesvirinae. Taken together, these results indicated that PRV strains were in the progress of evolution. This study has expanded the knowledge of genetic profiles of PRV strains.     

防治西洋参立枯病木霉菌株的筛选鉴定及其小区防治效果

摘要：利用平板对峙实验及小区试验,筛选高效防治西洋参立枯病的木霉菌株,并采用ITS（internal transcribed spacer)序列分析与形态学特征相结合的方法进行种类鉴定。结果表明,供试木霉菌株在PDA平板上对立枯丝核菌(R. solani)均有抑制作用,菌株HB20111效果最好,抑制率达99.33%;菌株HB20111对西洋参立枯病的小区效果显著,与其他处理间存在显著性差异(P<0.05);第1年拌种处理对西洋参立枯病的防治效果、单株参增重率及总参增重率分别达71.81%、33.54%和92.75%;第2年蘸根处理对西洋参立枯病的防治效果、单株参增重率及总参增重率分别达91.04%、158.06%和1 369.34%;菌株HB20111的分生孢子梗为典型的单侧分枝,其他形态学特征同深绿木霉(T. atroviride)一致,ITS序列与T. atroviride同源性达100.0%,将菌株HB20111鉴定为T. atroviride。HB20111生长速度快,产孢能力强,生防效果高,有望替代化学杀菌剂应用于西洋参种植中。     

Cloning and characterization of <Emphasis Type="Italic">UL</Emphasis>34 gene of pseudorabies virus HB isolate

In this study, China isolate HB of pseudorabies virus (PRV) was confirmed and genotypically characterized by amplifying and sequencing of partial UL34, a conservative gene involved in the egress of nucleocapsids from the nucleus, for phylogenetic analysis. The open reading frame (orf) of UL34 of PRV HB isolate is composed of 786 nucleotides, which encoded 262 amino acids. In addition, a potential transmembrane domain (241-260 aa) and 11 potential phosphorylation sites were also found in the UL34 of PRV HB isolate. Multiple amino acids alignment indicated that UL34 proteins of PRV strains derived from different geographic origins were highly conservative, but some mutations were also found. Phylogenetic analysis based on UL34 protein indicated that PRV HB strain was evolutionarily distinct from other recent China strains sequenced so far, forming a single clade within the phylogeny. Moreover, PRV HB isolate had close evolutionary relationship with BoHV-1 and BoHV-5 within the Alphaherpesvirinae. Taken together, these results indicated that PRV strains were in the progress of evolution. This study has expanded the knowledge of genetic profiles of PRV strains.     

抗稻瘟病几丁质酶产生菌的筛选及产酶条件的研究

稻瘟病具有传染快、危害大等特点,其微生物防治具有良好的发展前景.本实验采用以稻瘟病细胞壁为筛选培养基来筛选抗稻瘟病的几丁质酶产生菌,并对筛选到的优良茵株进行产酶条件优化.实验结果表明,从不同来源的土壤和植物器官中分离得到16株能检测到几丁质酶活性的几丁质酶产生茵,其中以H3和H9的产酶能力最强,具有良好的开发价值.     

辣椒疫霉菌拮抗细菌的筛选、鉴定及防效测定

从天津、河北、山东、山西、福建、河南及江苏等地蔬菜根际土壤中分离细菌1万余株,从中筛选出对辣椒疫霉菌Phytophthora capsici有明显拮抗作用的细菌802株,进一步用立枯丝核菌Rhizoctonia solani、番茄灰霉菌Botrytis cinerea及瓜果腐霉菌Pythium aphanidermatum为靶标筛选出对以上病原真菌有较好拮抗作用的菌株62株。对辣椒疫病防效温室盆栽实验结果表明,6个菌株防效在70%以上,其中菌株TB1340防效最高,达到82%。菌株TB1340生物学特性测定结果表明其具有较强的产生几丁质酶、纤维素酶和β-1,3-葡聚糖酶活性。经16S rRNA基因序列分子鉴定结合形态学观察,菌株TB1340被初步鉴定为芽孢杆菌Bacillus sp.。     

土壤中产a-淀粉酶菌株的分离和筛选

目的：从土壤中筛选产a-淀粉酶活力较强的菌株并对其酶学性质进行初步研究。方法：结合选择性分离培养,碘液检测法,酶活力测定和形态学检测等方法从土壤中分离出产酶活力较强的菌株,并对其所产仅一淀粉酶的最适反应温度和pH值进行研究。结果：共分离到4株能产a-淀粉酶的菌株,其中菌株4产酶活力最高。该菌株所产仅一淀粉酶最适反应温度为50～60℃,最适pH值为6．2。结论：分离到1株产a-淀粉酶酶活力较强的芽孢杆菌,所产仪一淀粉酶属于中温淀粉酶,最适pH值为6．2。     

对鸢尾新鲜根状茎发酵生香的微生物研究

对影响鸢尾新鲜根状茎发酵生香的微生物进行了初步探讨．从已陈化产香的鸢尾根状茎干片中分离到细菌９株，丝状真菌１４株．细菌中能产香的有２株，均属枯草芽孢杆菌（Ｂａｃｉｌ－ｌｕｓｓｕｂｔｉｌｉｓ），真菌中产香的只１株，为米根霉（Ｒｈｉｚｏｐｕｓｏｒｙｚａｅ）．实验证明，某些微生物对新鲜鸢尾根状茎的产香具有良好作用．经ＧＣ／ＭＳ分析表明，微生物发酵产物中含有鸢尾酮