壬基酚降解菌产酶条件的优化

通过单因素试验和正交试验确定了实验室保藏的壬基酚降解菌株沙雷氏菌(Serratiasp.LJ)的最佳产酶条件:以壬基酚、硫酸铵为碳源、氮源,培养基初始pH为6.8,培养温度为30℃,种子活化时间为24h,接种量为3%(体积分数)。在此条件下培养72h后,最高酶活力为1.314IU/mL,是优化前的1.77倍。     

黑曲霉固态发酵三七渣产淀粉酶的研究

以三七渣为基质,采用黑曲霉固态发酵产淀粉酶,考察了硫酸铵(氮源)添加量、三七渣粒径、固体培养基含水率、发酵温度、发酵时间、菌悬液接种量等因素对产淀粉酶效果的影响,并采用正交实验对发酵条件进行了优化。单因子实验结果表明,黑曲霉固态发酵三七渣产淀粉酶适宜的硫酸铵添加量为40～60mg/g(以干药渣计),最佳三七渣粒径为100目,最适固体培养基含水率为55%,最佳发酵温度为34℃,最适发酵时间为7d,最佳菌悬液接种量为10%(质量分数)。正交实验多重比较的结果表明,优化的发酵条件为:硫酸铵添加量50mg/g,发酵温度34℃,发酵时间5d,在此发酵条件下,黑曲霉固态发酵三七渣产生的淀粉酶的酶活可达84.15U/g(以湿物料计)。     

书虱伴生菌中纤维素酶产生菌的筛选、鉴定及最佳产酶发酵条件的优化

从以啃食树皮为生的书虱伴生菌中筛选得到5株产纤维素酶的菌株,编号分别为S2、S6、N10、N11和N12。结合菌株的形态学及16S r DNA序列分析等结果将这些菌株分别鉴定为Bacillus methylotrophicus,Streptomyces sp.,Pseudomans fluorescens,Bacillus sp.及Pseudomans sp.。分别使用单因素分析法和响应面分析法对纤维素酶活最高的S2菌株的产酶发酵条件进行了优化。单因素实验结果显示,B.methylotrophicus S2的最适产酶发酵条件为:0.01 g/m L的CMC碳源、10 g/L蛋白胨的氮源、48 h的发酵时间、28℃、初始pH为7.0,此时总纤维素酶活达到204.37 u/g。然后选取发酵时间、温度和初始pH作为3个因素,通过BBD实验,用响应面法对S2的发酵条件进行优化分析,最后得到一个拟合度良好的二次多项方程模型(R2=0.9948)。方差分析结果显示,发酵温度与培养基初始pH之间的交互作用极显著。响应面分析优化后的反应体系为:温度24℃、初始pH为7.7、发酵59 h,测得酶活力为303.18 u/g,比优化前单因素最佳纤维素酶活力提高45%。     

谷氨酸发酵废菌体的水解与替代酵母膏生产多聚谷氨酸

为研究谷氨酸发酵废菌体水解液用作培养基氮源的可行性,以蛋白质水解度和溶解度为衡量指标,采用化学、生物、物理以及这几种方法的联合处理,通过正交优化实验,得出了菌体蛋白水解的最佳工艺条件为化学-生物联合处理,在盐酸浓度3 mol·L~(-1)、温度110℃、料液比1:0.5、水解12 h后使用酸性蛋白酶酶解,调节料液pH为4、酶底比2.5%、温度55℃、酶解8 h,水解完成后蛋白质水解度达到47.18%,溶解度达到94.09%。在总氮相等的情况下,将水解液和酵母膏以不同的比例组合,替代正常发酵产γ-聚谷氨酸过程的氮源。结果表明谷氨酸发酵废菌体经过最佳工艺水解后,其水解液可以用作发酵产γ-聚谷氨酸的氮源。     

固态发酵法制备优势硝化菌固定化产品的工艺优化

为解决高效菌在工程应用中扩大培养、保存和接种等瓶颈,研究固态发酵法制备焦化废水优势硝化菌固定化产品.以TTC脱氢酶活性法作为固体发酵生物量测定指标,通过液态发酵单因素实验初步确定硝化菌发酵条件,再用响应面法对其固态发酵条件进行优化.结果表明,TTC脱氢酶活性法适合作为表征生物质载体固定化产品生物量的测定方法,不受载体和培养时间的影响,能较好地指示干燥前固态发酵生物量的变化.固态发酵主要影响参数依次为发酵时间、发酵温度、接种量和含水率.发酵时间的最佳值为47 h,发酵温度最佳为29.4℃,接种量的最佳值为9.9 mL/200 mL,最佳含水率为48.9％时,固态发酵固定化产品酶活达到17.091 μg TF/(g·h).与液态发酵参数相比其发酵温度、接种量最佳值接近,但发酵时间由1d延长至2d,产品酶活持平.     

不同存放时间太湖蓝藻产沼气潜力

对不同存放时间蓝藻(新鲜蓝藻、存放3个月、6个月和9个月)进行厌氧消化处置,分析反应过程中沼气量、代谢产物和酶活等变化,获得各自的产沼气潜力。研究结果表明,存放6个月蓝藻组的产沼气率最大,为120.5 mL/g TS,是新鲜蓝藻组的1.29倍。Gompertz模型拟合表明,随着存放时间的延长,最大产沼气量(Pmax)先增大后减小,最大比产气率一直增加。存放3个月蓝藻组的VFA在第6天达到最大,为1 575.4 mg/L,而存放9个月蓝藻组的VFA在第2天达到最大,为1 116.5 mg/L。存放3个月蓝藻组的最大中性蛋白酶酶活为1 203.4μg/(g TS.h),比新鲜蓝藻组提高212.8μg/(gTS.h)。脱氢酶酶活随着蓝藻存放时间的增加呈现先增加后减小的趋势。     

PVA降解混合菌系生长和产酶的营养条件分析

研究碳源、氮源以及不同金属离子对PVA降解混合菌系生长和产酶的影响。结果表明,混合菌系生长过程中无机氮源优于有机氮源;补充碳源可促进混合菌系的生长,但酶活比不添加时稍有降低。金属离子对酶活影响的正交实验表明Mg2＋浓度影响最大,其次是Fe2＋和K＋浓度,Ca2＋、Mn2＋、Zn2＋和Cu2＋浓度影响较小;最佳浓度分别为Mg2＋0.05 g/L、Fe2＋0.04 g/L、K＋2 g/L和Ca2＋0.05 g/L。     

玉米浸泡液制备苏云金杆菌生物杀虫剂的影响因素研究

以玉米淀粉生产过程中的浸泡液为培养基,摇瓶发酵培养苏云金杆菌生物杀虫剂,通过一系列单因子试验,考察了不同培养条件(种子液的种龄、接种量、浸泡液的含固率、初始pH值、摇床转速、发酵温度及发酵时间)对苏云金杆菌在玉米浸泡液中的生长(菌数增长与芽孢形成)以及发酵液的生物毒效的影响.研究表明,在最佳摇瓶培养条件(种子液种龄10 h,接种量2%,浸泡液含固量3%,初始pH值7.0～7.5,摇床转速200 r/min,发酵温度30℃)下发酵48 h,活菌数和活芽孢数分别可达到7.9×108 CFU/mL和5.5×108 CFU/mL,毒力效价为698.0 IU/μL.本试验可为生物农药的工业化生产提供实用参数.     

PVA降解菌的复配组合及其紫外诱变处理的研究

对已筛选出的4株PVA降解菌进行不同的组合后测定PVA降解率,发现Z5与Z8菌株混合后在1周内对PVA的去除率达到了所有组合中的最高值52.07%,比单菌株提高了约22%。采用紫外照射的方法对混合菌进行诱变并利用正交实验研究其特性,结果表明,诱变后的混合菌在1周内降解了91.46%的PVA,并且碳源浓度对酶活的影响最为显著。当pH为7,碳源、氮源含量分别为1.5 g/L、0.6 g/L,NaC l浓度为1.5 g/L时,诱变菌株处于最佳生长状态。     

海带渣废弃物资源化利用以及多功能菌肥固体发酵条件的优化

海带渣是海带加工过程中主要固体废弃物。本实验利用海带渣对生防菌株JGA2(-5)27-2进行固体发酵。实验对不同发酵条件下发酵产物对植物病原真菌寄生曲霉的抑制活性与对发酵产植物激素6-糠氨基嘌呤的能力进行测定。通过单因素实验和响应面实验优化了发酵条件:蔗糖为最优的碳源;硝酸盐为最优的氮源;蔗糖浓度、KNO3浓度、最佳的培养时间、含水量、培养温度、接种量和pH值分别为2.091%、0.507%、8.24 d、60%、35℃、4%和自然pH值。在发酵过程中以添加KNO3的形式向发酵菌肥中加入K+离子。实验证明,利用生防菌株JGA2(-5)27-2进行海带渣的固体发酵生产多功能微生物肥料具有可行性,并为海带渣的资源化在农业领域的利用提供一定参考。     

氮源对克雷伯氏菌NⅢ2分泌絮凝剂特性的影响

实验研究了蔗糖为碳源,硝酸钠、脲、蛋白胨、硫酸铵和氯化铵等氮源对NⅢ2发酵产絮凝剂的影响。结果表明,发酵液起始pH值为7.50,以硝酸钠为氮源,发酵液pH会上升,升至7.60～8.34时,NⅢ2菌株开始大量分泌微生物絮凝剂,发酵72 h,产量可达7.5 g/L,该产量是目前报道的克雷伯氏菌产絮凝剂的最高值。脲为氮源,pH则下降,降至5.04～6.49时,大量分泌絮凝剂,发酵72 h产量达5.2 g/L。蛋白胨、氯化铵和硫酸铵等为氮源时,pH下降十分明显,pH小于3.71时有絮凝剂分泌,发酵72 h产量约2.0 g/L或更小。以硝酸钠和脲为氮源时,发酵液中有黄色物质分泌,该黄色物质出现或黄色逐渐加深,是NⅢ2菌高产絮凝剂的标志。除硫酸铵外,其他氮源发酵所产絮凝剂为O-糖蛋白。当以硝酸钠、脲、蛋白胨、硫酸铵和氯化铵为氮源时,絮凝剂中蛋白的含量分别为9.55%、33.28%、19.39%、13.81%和15.51%,且蛋白含量越高,絮凝剂活性越大。     

生物浸取电解锰渣中锰的研究

为了评价生物法浸取电解锰渣中锰的可行性,利用锰渣土壤中筛选出的2种锰抗性强的微生物Serratia sp.和Fusarium sp.浸取电解锰渣中的锰,并采用优化的BCR（European Community Bureau of Reference）连续萃取方案对浸取前后的金属锰进行形态分析,研究其浸出率和浸取前后锰的形态变化特征。同时考察了3种萃取剂EDTA、HNO₃和CaCl₂对锰的萃取效率及萃取后金属锰的形态变化。研究结果表明,Serratia sp.和Fusarium sp.对锰都表现出一定的浸取能力,Fusarium sp.的浸取能力尤为显著,3 d后锰浸出率达到56.5%,为锰污染的微生物治理和资源化利用提供科学依据。3种萃取剂对锰的浸取效果为EDTA>HNO₃>CaCl₂,平均萃取效率依次为50.0%、28.8%和21.2%。浸取前后,酸溶解态锰所占比例变化较显著,说明酸溶解态锰是比较容易浸取的形态。     

Influence of fenamidone, indoxacarb, pyraclostrobin, and deltamethrin on the population of natural yeast microflora during winemaking of two sardinian grape cultivars

The influence of fenamidone ((S)-1-anilino-4-methyl-2-methylthio-4-phenylimidazolin-5-one), pyraclostrobin (methyl 2-[1-(4-chlorophenyl)pyrazol-3-yloxymethyl]-N-methoxycarbanilate), indoxacarb (methyl 7-Chloro-2,5-dihydro-2-[[(methoxycarbonyl) [4- (trifluoromethoxy) phenyl] amino] carbonyl] indeno[1,2-e][1,3,4] oxadiazine-4a(3H)-carboxylate), and deltamethrin ([cyano-[3-(phenoxy)phenyl]methyl] 3-(2,2-dibromoethenyl)-2,2-dimethylcyclopropane-1-carboxylate) on spontaneous fermentation carried out by natural yeast grapes microflora, was studied during the wine-making process. Aliquots of pesticide standard solutions were added to the grapes before crushing, to reach a concentration equal or half the maximum residue limit (MRL). Vinifications were performed, with maceration (R), or without maceration (W). During the wine-making process, samples were taken at the beginning (one hour after grapes crushing), at the middle and at the end of the spontaneous fermentation process. At half the MRL concentration, deltamethrin affected Pichia sp. population with a decrease of almost 50 %, while fenamidone decreased Candida sp., Candida stellata at 83, and 36%, respectively. Metschnikowia pulcherrima population decreased in all samples when compared to the control. Experiments at MRL levels showed a strong reduction for all non-Saccharomyces yeast species, when grapes had been treated with pyraclostrobin, fenamidone, and deltamethrine, except for Candida sp. which was found to have been affected only by fenamidone residues. Growth zone inhibition test showed only an in vitro activity of pyraclostrobin over Kloeckera spp., C. stellata, and M. pulcherrima. Microvinification experiments produced wines with no differences concerning S. cerevisiae population as well as production of ethanol and residual sugars. Experiments showed that at the end of the fermentation process pesticides were adsorbed by the lees and grape skins, and no pesticides residue was detectable in wine.     

Microbial decolorization of synthetic dyes and reactive dyes of industrial effluents by using a novel fungus Aspergillus proliferans

A decolorizing fungal strain was isolated and identified by the morphology and genotypic characterization as Aspergillus proliferans. The effect of A. proliferans on decolorization of synthetic dyes (70 mg ml(-1)) and colored effluent was evaluated in liquid culture medium. A. proliferans expressed their effective decolorization activity in effectual decolorization of synthetic dyes and industrial effluent. Synthetic dyes were decolorized by 76 to 89% within 6 days of treatment and 73.5% of color was removed in industrial effluent within 8 days. The addition of optimum carbon and nitrogen sources were effectively stimulated the decolorization activity. The high concentration of glucose repressed the decolorization activity and supplementation of yeast extract has significantly enhanced the effluent decolorization at p < 0.05. Laccase enzyme was isolated from liquid state fermentation, which showed significant enzyme activity (10,200 Uml(-1)) at p < 0.005. The crude enzyme decolorizes the dyes aniline blue and congo red in 14 hours (40.9 to 70%) and the effluent in 14 hours (88.6%). Moreover, the culture free supernatant without the fungal biomass has also effectively decolorized the effluent and synthetic dyes. The fungi Aspergillus proliferans was used not only for decolorization but also for better bioremediation of industrial effluent.     

Lysis of cyanobacteria with volatile organic compounds

One of bacteria collected from Lake Sagami, Japan, Brevibacillus sp., was found to have a lytic activity of cyanobacteria, but did not produce active compounds. Instead, the co-culturing of Microcystis with the Brevibacillus sp. enhanced the production of two volatile compounds, beta-cyclocitral and 3-methyl-1-butanol, and the former had a characteristic lytic activity. It was confirmed that these volatile compounds were derived from the cyanobacteria themselves. beta-Ionone, geosmin and 2-methylisoborneol derived from cyanobacteria and similar volatile compounds, terpenoids, produced by plants also had a lytic activity. The minimum inhibitory concentration values of the cyanobacterial metabolites were estimated to be higher than those of compounds from plants except for a few compounds. Among them, beta-cyclocitral only produced a characteristic color change of culture broth from green to blue. This color change is similar to the phenomenon observed when a sudden decline in growth of cyanobacteria begins in a natural environment.     

培养基对菌产微生物絮凝剂的影响研究

研究了培养基成分种类及其用量对菌产微生物絮凝剂的影响。结果表明,较高的C/N比对菌产絮凝剂有利,碳源中的蔗糖是菌株Aeromonassp.N11产絮凝剂的良好碳源,氮源以采用复合氮源为佳。培养基中加入酵母膏有利于絮凝剂的产生。NaCl能够促进所产絮凝剂的絮凝活性,碳源蔗糖和氮源脲对菌产絮凝剂的影响最大。利用正交实验得出产絮凝剂培养基的最佳配比为蔗糖20g/L,酵母膏08g/L,脲05g/L,硫酸胺05g/L,NaCl7g/L。     

Arsenosugars and other arsenic compounds in littoral zone algae from the Adriatic Sea

In 10 different marine algae from the littoral zone (found between the highest and lowest tide marks on the seashore) arsenic compounds were determined by means of a high-performance liquid chromatography (anion and cation exchange)-UV photochemical digestion-hydride generation-atomic fluorescence spectrometry (HPLC-UV-HGAFS) system. Samples (Ceramium sp., Cystoseira barbata, Enteromorpha sp., Fucus virsoides, two different species of Gelidium, Padina pavonica, Polisyphonia sp. and Ulva rigida) were collected along the Adriatic Sea coast of Slovenia. The total arsenic content of the algal samples, as determined by ICP-MS, ranged from 1.35 to 28.1 microg g(-1) (fresh weight). In all algae but two, the most abundant arsenic species found were arsenosugars with minor amounts of other arsenic compounds. Cystoseira barbata and Ceramium sp. contained high amounts of mainly inorganic arsenic. A small quantity of arsenobetaine was detected in most of the investigated Adriatic algae, which probably originates from mesofauna attached to the algae in their natural habitat.     

4种固定化藻类对污水中氮的净化能力研究

取培养至对数末期的藻,采用海藻酸钙凝胶包埋固定,对人工污水进行静态模拟净化试验,研究了蛋白核小球藻、鱼腥藻、双对栅藻和突变衣藻4种藻在固定和悬浮状态下,对污水中的氨氮和硝酸氮的净化效率以及藻类的生长特性。结果表明：固定化藻细胞比悬浮态藻细胞具有生长更趋于稳定、藻类的活性保持时间更长的优势。4种藻类中,小球藻和鱼腥藻在污水中的生长状况更好,较适宜采用海藻酸钙凝胶包埋固定化技术。实验第5 d时,固定化小球藻、鱼腥藻、双对栅藻和衣藻对NH₃-N去除率分别为91.9%、84.8%、68.3%和51.2%;对NO^-₃-N的去除率分别为85.1%、100%、96.9%和65.9%。固定化小球藻对NH₃-N的去除效果最好,而固定化鱼腥藻对NO^-₃-N的去除效果最好。因此,小球藻和鱼腥藻更适用于去除污水中的氮,具有很好的应用前景。     

Effects of cadmium on sediment processing on members of the Capitella species-complex

The effect of cadmium on sediment processing by laboratory-cultured specimens of Capitella sp I and Capitella sp B was investigated. Specimens were exposed to 0, 60, 90 and 140 microg Cd g(-1) dry weight sediment for a 10 day period (two census days) to observe effects of cadmium on their feeding activity. Under unstressed conditions, Capitella sp I specimens were larger, had higher biomass and produced more fecal pellets than Capitella sp B. Cadmium concentrations had no effect on pellet production and body mass of the studied populations. However, cadmium exposure time significantly affected Capitella sp B pellet production and body mass of both populations following different trends: after 10 days of exposure, the average processed sediment per unit worm mass decreased in Capitella sp I but increased in Capitella sp B. The latter population did not reduce its feeding activity in the presence of cadmium maybe related to the fact that it is derived from a highly polluted environment. This study has shown an unimpaired response of Capitella sp I and Capitella sp B to cadmium in their sediment processing activity, which indicates the potential importance of this species complex in the trophic transfer of metals from sediments to other marine organisms.