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1.
从长期施用多菌灵农药的土壤中,通过富集筛选,获得1株新的多菌灵高效降解菌株。通过生理生化实验和16SrDNA序列同源性分析鉴定该菌株,应用高效液相色谱法对纯培养条件下菌株的降解特性和粗酶提取液的降解性能进行了分析。结果表明,筛选所获得的菌株与Raoultella菌属的亲缘关系最近,将其命名为Raoultellasp.MBC,该菌株能在以多菌灵为唯-碳源的无机盐培养基中生长;25℃、pH7.0、200r·min。的最适生长条件下避光振荡培养72h,多菌灵的降解率达到100%;在最适培养条件下外加氮源和碳源在培养后期均可以提高多菌灵的降解率,外加氮源对多菌灵的降解效果优于外加碳源;该菌体的粗酶提取液具有降解多菌灵活性,且多菌灵降解酶为诱导酶。研究结果为多菌灵污染土壤的生物修复和酶修复提供了材料和理论依据。  相似文献   

2.
以魔芋生粉为唯一碳源的筛选培养基,从魔芋废渣中分离筛选出一株可以降解魔芋生粉的细菌菌株,编号为LS-6。根据16SrDNA序列和生理生化特性,将该菌株鉴定为枯草芽孢杆菌Bacillus subtilis。胞内外葡甘露聚糖酶酶活分析结果表明,LS-6产生的葡甘聚糖酶为胞外酶。LS-6产生葡甘聚糖酶的最佳培养基组分和条件是:1%魔芋生粉,0.5%酵母粉,0.5%NaCl,培养基初始pH为6.0,最佳发酵条件是25℃,摇床转速200r/min,培养48h。酶解产物的高效液相色谱分析结果表明,LS-6粗酶液的酶解产物主要为葡萄糖和甘露糖。LS-6的分离为进一步克隆常温葡甘聚糖酶基因和产酶工程菌的构建奠定了基础。  相似文献   

3.
集约化畜禽养殖场产生的沼液通常就地回用,在循环利用有机物的同时也会带来类固醇雌激素(Steroid Estrogens,SEs)的累积及污染。为降低沼灌后SEs对水土环境的污染风险,该研究采用富集和纯化培养法,对西南地区某奶牛养殖场沼灌区土壤中雌激素降解菌进行分离及筛选,获得一株利用17β-雌二醇(17β-E2)为唯一碳源生长繁殖的降解菌。通过16S rDNA 基因序列进行同源性比对以确定种属,并研究其降解特性。分别研究了菌株在不同温度、pH值、底物浓度三种单因素条件下的降解特性,然后利用三因素三水平正交试验继续优化菌株最适降解条件。结果表明:分离出的优势菌为生丝微菌属(Hyphomicrobium sp.),命名为Hyphomicrobium sp.SS-1,该菌株在10~40 ℃、pH值为5~9、底物浓度为1~10 mg/L的条件下,均能不同程度降解17β-E2。其中菌株在温度为30 ℃、pH值为7、底物浓度5 mg/L的条件下,培养7 d对17β-E2的降解率可达71%,并伴随毒性低于E2的降解产物E1和E3生成,总雌激素去除率为56.8%。正交试验结果显示,各因素对菌株降解能力的影响顺序从小到大为:底物浓度、温度、pH值,且都为显著影响(P<0.05);菌株最适降解条件为温度35 ℃、pH值为7、底物浓度5 mg/L,该条件下培养7 d,菌株对17β-E2的降解率可达97.09%。研究结果可为复杂基质环境中微生物降解SEs提供优质菌种资源,并为沼液灌溉区土壤的雌激素污染修复提供有效途径。  相似文献   

4.
  【目的】  海藻 (海带) 降解物对作物具有抗逆促生作用,研究海带菌解和酶解的最佳工艺条件,并比较两个工艺生产的降解产物对菜心的抗逆效果。  【方法】  采用单因素和正交试验,探究不同的底物浓度、接种量、溶液pH、降解温度分别对微泡菌Microbulbifer sp. SH-1和海藻酸裂解酶AlgSH7制备海藻降解产物的影响。同时,借助光学显微镜,研究SH-1菌和AlgSH7酶降解过程中海带细胞形态变化。并在此基础上,利用盆栽试验,研究海带菌解产物、酶解产物对菜心非生物胁迫抗性的影响。  【结果】  海藻酸裂解酶AlgSH7降解海带的最佳工艺条件是底物浓度2%,酶添加量6%,降解体系初始pH 8.5,温度44℃;SH-1菌株降解海带的最佳工艺条件是底物浓度2.5%,接种量1.5%,降解体系初始pH 7.5,温度32℃。与菌解产物相比,酶解产物中海藻酸、总糖、还原糖含量分别提高141.8%、57.6%、150.5%,而褐藻多酚、甘露醇、甜菜碱含量分别下降35.3%、60.6%、62.6%,但菌解工艺的产率比酶解工艺高9.3%。细胞形态观察结果表明,海带原始细胞排列紧密,形状规则饱满,随菌解或酶解时间的延长,海带细胞可视面积变小,细胞之间的间距增大。降解24 h时,酶解海带细胞可视面积仅为菌解的77.2%。在盆栽试验中,中度干旱条件下 (土壤相对含水量50%),酶解液灌根处理的菜心生物量比菌解液处理提高13.7%;酶解液配施水溶肥 (NPK, N:P2O5:K2O=110:50:60) 处理的菜心生物量比菌解液配施NPK处理明显提高10.6%。淹水条件下 (淹水层深度2 cm),酶解液处理的菜心生物量比菌解液处理降低7.1%,但均显著高于对照处理;酶解液配施NPK处理的菜心生物量比菌解液配施NPK处理提高5.6%。盐胁迫条件下 (土壤中NaCl 含量24 g/kg),海带酶解液处理的菜心生物量比菌解液处理有增加趋势,但增加不显著。  【结论】  海带酶解产物中海藻酸、总糖、还原糖含量高于菌解产物,而褐藻多酚、甘露醇、甜菜碱含量低于菌解产物。菌解海带的产率高于酶解海带的产率。逆境胁迫条件下,海带酶解液配施NPK处理的菜心生物量高于菌解液配施NPK处理,海带酶解液对农作物抗逆促生作用优于菌解产物。  相似文献   

5.
陈晶晶  陶少强  夏强  王雅楠  秦冰  朱林 《土壤》2014,46(2):302-307
利用小麦/玉米秸秆还田土壤样品,通过富集培养和刚果红平板染色法筛选分离出纤维素降解细菌XWS-12;对分离的菌株进行16S rRNA基因序列系统发育分析,初步鉴定为伯克氏菌属(Burkholderia),定名为Burkholderia sp.XWS-12。以玉米秸秆和麸皮为碳源,研究了氮源、发酵时间、初始发酵温度、培养基初始pH等条件对该伯克氏菌产纤维素酶的影响。结果显示,该菌株产纤维素酶最适氮源为硝酸钠,培养时间为60 h,培养温度为37℃,培养基初始pH为4,该菌株的CMC酶活力最高,可达25 U/ml。其粗酶液的最适反应温度为50℃,最适反应pH为5,在pH 4~8的范围内酶活力较稳定。粗酶液的热稳定较差,当温度超过50℃时,该酶活力显著下降;当温度为50℃时,保温1 h,该酶活力损失53%。  相似文献   

6.
运用恒温培养法研究了菲在不同土壤中的降解动态,结果表明未灭菌土壤中菲半衰期仅5.1d,而灭菌土壤中半衰期为前者4倍,可见土壤微生物发挥了重要作用;从试验土壤中分离到一株菲高效降解菌,经16SrDNA鉴定为产碱杆菌;优势菌最高可以耐受500mgL-1左右的菲,降解菲的最适温度25℃和pH为7,土壤接种优势菌后有助于加快菲的降解;从该优势菌中提取的粗酶液米氏常数为50.30nmolml-1,最大降解速率为171.33nmolmg-1min-1。  相似文献   

7.
合理降解烤后烟叶中的淀粉和纤维素含量已成为提高烟叶质量的关键工艺之一。本研究对分离自烤烟叶面的产淀粉酶细菌菌株解淀粉芽孢杆菌(Bacillus amyloliquefaciens A1)和产纤维素酶细菌菌株短小芽孢杆菌(B.pumilus C1)进行发酵培养,提取粗酶液,检测酶活性并应用于初烤烟叶上。通过设计不同酶量、作用时间、相对湿度和温度的方法考察这两种酶制剂对烟叶(Nicotiana tabacum K326)中淀粉和纤维素的降解效果。结果表明,菌株A1主要产α-淀粉酶,酶活力为7×105U/mL,菌株C1以产内切酶为主,酶活力为6×103U/mL;在温度40℃、湿度70%条件下作用96h,用菌株A1所产淀粉酶制剂4×107U/kg处理后的烟叶总糖增加率为12.78%,还原糖增加率为12.03%,菌株C1所产纤维素酶制剂4×105U/kg处理后的烟叶总糖增加率为13.87%,还原糖增加率为18.07%。研究结果提示,利用从烟叶表面分离的产酶菌株进行发酵生产的酶制剂可降解初烤烟叶中淀粉和纤维素,提高还原糖含量,可望在烟叶加工过程中得到应用。  相似文献   

8.
王会娟  刘阳  邢福国 《核农学报》2012,26(7):1025-1030
通过选择培养基平板培养法和液体发酵培养法筛选得到2株高产漆酶的平菇菌株P1和P2,并对平菇菌株产漆酶的培养基进行筛选,得到产漆酶的最适培养基为最低盐MSM培养基。菌株P1不仅产漆酶能力最高,而且降解黄曲霉毒素的能力也最好。在MSM培养基中培养10d时,产漆酶量高达769.44U/L,在800μl的反应体系中,790μl粗酶液可以将1000ng黄曲霉毒素B1降解到222.62ng,降解率为77.74%,并且平菇粗酶液降解黄曲霉毒素B1的能力与其中漆酶的含量呈一定的正相关性。  相似文献   

9.
机械活化玉米淀粉的微生物降解性能   总被引:4,自引:1,他引:3  
为了提高玉米淀粉的微生物降解反应活性,采用搅拌球磨机对玉米淀粉进行机械活化,以活化时间为60 min的玉米淀粉和原淀粉为原料,酒曲为降解试剂进行微生物降解反应,并以降解产物的葡萄糖值(Dextrose Equivalent,DE)为评价指标,分别研究了糊化温度、pH值、淀粉浓度、降解时间、降解温度、酒曲培养液用量等因素对降解产物中葡萄糖值的影响,并采用扫描电镜对淀粉降解过程中的颗粒进行形貌观察。结果表明,机械活化预处理能提高玉米淀粉微生物降解反应液中葡萄糖的含量,且活化淀粉未经糊化就能直接被微生物降解,降解60 min时的DE值为36.76%,与原淀粉相比,提高了27.80个百分点。说明机械活化作用破坏玉米淀粉紧密的颗粒表面和结晶结构,有效地提高了微生物降解反应活性。  相似文献   

10.
为实现砂姜黑土地区秸秆资源化利用以及作物增产,本研究从该区小麦-玉米轮作土壤中筛选具有促生功能的纤维素降解菌株。通过羧甲基纤维素酶活(CMC)与吲哚乙酸(IAA)分泌量测定,筛选出1株具有高效降解纤维素的促生菌株。经形态学和分子生物学鉴定,该菌株为普沙根瘤菌(Rhizobium pusense),命名为X2。小麦秸秆降解和玉米盆栽初步试验结果显示,X2菌株经液态发酵15d后小麦秸秆降解率为16.1%,较对照显著提升65.4%;接种该菌株后盆栽土壤中碱解氮含量较对照显著提高了72.7%,植株根系的平均直径、表面积以及地上部干物质量显著增加了22.0%、28.6%和33.3%。为进一步提高该菌株的活性,采用单因素试验对菌株的培养条件进行优化,结果表明,X2菌株在装液量为25 mL/250 mL、氮源为酵母粉、pH为6.0时生长和产IAA能力最优,产酶的最适pH为5.0。综上,所筛菌株X2具有秸秆降解和玉米促生能力,可在砂姜黑土地区为秸秆资源化利用提供新的微生物资源。  相似文献   

11.
To develop safe and cheap thrombolytic agents, a fibrinolytic enzyme productive strain of LSSE-62 was isolated from Chinese soybean paste. This strain was identified as Bacillus amyloliquefaciens by 16S rDNA sequence analysis. Nucleotide and amino acid sequence analysis showed that this fibrinolytic enzyme was identical to subtilisin DJ-4. Chickpeas were used as the substrate for fibrinolytic enzyme production from B. amyloliquefaciens in solid-state fermentation. Under the optimized conditions (34 °C and 50% initial moisture content), the fibrinolytic activity of fermented chickpeas reached 39.28 fibrin degradation units (FU)/g. Additionally, the fermented chickpeas showed anticoagulant activity, and the purified anticoagulant component showed higher anticoagulant activity than heparin sodium. After fermentation, the total phenolic and total flavonoid contents increased by 222 and 71%, respectively, and then the antioxidant activities were improved significantly. This study provided a novel method for the preparation of multifunctional food of chickpeas or raw materials for the preparation of functional food additives and potential drugs.  相似文献   

12.
本文以3个亲缘关系较远的高产木薯品种(SC124,SC8和Arg7)为材料,对块根发育过程中的AGPase活性和淀粉含量进行了测定,并利用AGPase同工酶电泳技术(非变性聚丙烯酰胺凝胶电泳)对AGPase酶活性较高时期同工酶位点进行检测,旨在分析木薯块根AGPase同工酶位点与AGPase酶活性和淀粉积累的关系。结果表明:木薯块根发育过程中AGPase酶活性和淀粉含量呈极显著正相关(R=0.80212),Arg7的AGPase酶活性和淀粉含量在块根发育都显著高于SC124和SC8。同工酶分析表明,木薯至少有6个同工酶位点(AGPa,AGPb,AGPc,AGPd,AGPe和AGPf),且品种间具有多态性。其中,3个品种共有的4个同工酶位点分别为AGPa、AGPc、AGPd和AGPf,AGPe位点只在Arg7中出现,初步判断同工酶位点AGPe可能与其较高的酶活性和淀粉含量有关。本研究为系统研究木薯品种间AGPase酶活性与淀粉积累的关系提供了参考。  相似文献   

13.
One of the objectives of the cassava-breeding project at CIAT is the identification of clones with special root quality characteristics. A large number of self-pollinations have been made in search of useful recessive traits. During 2006 harvests an S1 plant produced roots that stained brownish-red when treated with an iodine solution, suggesting that it had lower-than-normal levels of amylose in its starch. Colorimetric and DSC measurements indicated low levels (3.4%) and an absence of amylose in the starch, respectively. SDS-PAGE demonstrated the absence of GBSS enzyme in the starch from these roots. Pasting behavior was analyzed with a rapid visco-analyzer and resulted in larger values for peak viscosity, gel breakdown, and setback in the mutant compared with normal cassava starch. Solubility was considerably reduced, while the swelling index and volume fraction of the dispersed phase were higher in the mutant. No change in starch granule size or shape was observed. This is the first report of a natural mutation in cassava that drastically reduces amylose content in root starch.  相似文献   

14.
一株耐辐射考克氏菌的分离与鉴定   总被引:2,自引:2,他引:0  
唐然  袁梦龙  吴菁  陈明  张维  林敏 《核农学报》2010,24(2):276-280
从10kGy60Coγ射线辐照处理的新疆戈壁土壤中分离到一株耐辐射微生物I-7R,该菌菌落为橙红色革兰氏阳性球菌。最适生长温度为30℃,(G+C)mol%含量为67.5%,不具有氨苄青霉素、氯霉素、四环素、卡那霉素、潮霉素、利福平及壮观霉素等抗性。UV辐射生存曲线显示,I-7R菌株与大肠杆菌K-12相比,具有较强的紫外辐射抗性。16S rDNA序列比较分析表明I-7R菌株与Kocuria rosea、K.erythromyxa以及K.polaris同源性达到99%。结合I-7R菌株生理生化试验结果,该株菌归属于考克氏菌属(Kocuria),并与Kocuria rosea最为相近,暂命名为K.rosea I-7R。  相似文献   

15.
The observation that the bitterest peptides from casein hydrolysates contain several proline residues led us to hypothesize that a proline-specific protease would be instrumental in debittering such peptides. To identify the desired proline-specific activity, a microbiological screening was carried out in which the chromogenic peptide benzyloxycarbonyl-glycine-proline-p-nitroanilide (Z-Gly-Pro-pNA) was used as the substrate. An Aspergillus niger (A. niger) strain was identified that produces an extracellular proline-specific protease with an acidic pH optimum. On the basis of sequence similarities, we conclude that the A. niger-derived enzyme probably belongs to the S28 family of clan SC of serine proteases rather than the S9 family to which prolyl oligopeptidases belong. Incubating the overexpressed and purified enzyme with bitter casein hydrolysates showed a major debittering effect. Reversed phase HPLC analysis revealed that this debittering effect is accompanied by a significant reduction of the number of hydrophobic peptides present.  相似文献   

16.
Four α‐amylases and two glucoamylases from various sources, in eight combinations, were used to study the synergistic hydrolysis of crude corn starch at various temperatures. At 40 and 50°C, the combinations containing Rhizopus mold glucoamylase enhanced hydrolysis of corn starch compared wth that obtained with the combinations from Aspergillus niger. At 60°C, Rhizopus mold combinations gave low reaction yields as the enzyme was inactivated. The differences observed between α‐amylases are smaller, with the exception of Bacillus licheniformis α‐amylase, which presented more than twice the productivity of the other α‐amylases, at all temperatures. In terms of substrate conversion at 5 hr of hydrolysis, the combination of B. licheniformis α‐amylase with Rhizopus mold glucoamylase at 50°C presents 76% substrate conversion, whereas, with all the other combinations, starch conversion was 13–73%. HPLC analysis of the reaction products obtained at 50°C showed that the main product of corn starch hydrolysis was glucose at 85–100%. Further experiments showed that A. niger glucoamylase and B. licheniformis α‐amylase were the only enzymes that retained their initial activity after incubation at the temperatures studied.  相似文献   

17.
本研究从广西花坪自然保护区采集的土壤中筛选获得了一株产糖化酶丝状真菌菌株57-45,通过形态学观察和真菌内转录间隔区(internal transcribed spacer,ITS)序列比对分析,将其初步鉴定为曲霉属(Aspergillus sp.)的一个种。纯化真菌57-45所产的一种胞外糖化酶经过硫酸铵分级沉淀、疏水层析和阴离子交换层析三步蛋白质纯化步骤,得到在SDS-PAGE上约60kD的单一蛋白质条带,薄层层析分析表明该纯化的蛋白质水解可溶性淀粉的产物只有葡萄糖,证明该纯化的蛋白质为糖化酶。纯化的糖化酶Km值为1.9mg/mL,Vmax为4148μmol/(min·mg),最适作用pH5.5,最适作用温度50℃,在同步糖化发酵中有应用的潜力。金属离子Fe3+、Zn2+、Cu2+对酶活有较强的抑制作用,EDTA对酶活有较强的促进作用。本文结果将为进一步研究曲霉糖化酶的酶学特性提供新的材料。  相似文献   

18.
The hydrolysis of raw starch catalyzed by glucoamylase has been studied with starch granules of different sizes by use of an amperometric glucose sensor by which the direct and continuous observation of the concentration of glucose can be achieved even in a thick raw starch suspension. The initial rate of the enzymatic hydrolysis in the raw starch suspension increased with increasing concentration of the enzyme to approach a saturation value and was proportional to the amount of substrate. Also, the rate was proportional to the specific surface area of the substrate. The experimental results can be explained well by the rate equations derived from a three-step mechanism, which consists of adsorption of the free enzyme onto the surface of the substrate, reaction of the adsorbed enzyme with the substrate, and liberation of the product.  相似文献   

19.
从农药厂废水中分离到能够降解多种拟除虫菊酯的复合菌群JZ-1,对该菌群降解特性研究结果表明,最佳降解条件为pH7、温度30℃;在最佳条件下培养15d,对100mg·L-1甲氰菊酯、氟氯氰菊酯、氯氰菊酯的降解率分别为53.27%、33.36%、41.39%。对该菌群的16S rDNA进行扩增和RFLP及测序分析结果表明,该菌群含有丰富的细菌资源,该菌群的优势种群包括红假单胞菌属(Rhodopseudomonas)和紫单孢菌科细菌(Porphyromonadaceae bacterium)。该研究为拟除虫菊酯的降解菌资源的挖掘和利用提供了理论参考。  相似文献   

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