采油废水模拟处理系统中酵母菌群落结构动态初步解析

本研究建立好氧活性污泥的模拟体系处理经过物化预处理的采油废水, 控制温度等条件接近实际运行体系的情况下, 考察系统COD去除率和污泥体系中酵母菌多样性变化。结果显示系统稳定后COD去除率在70%以上; 利用非培养的方法考察了污泥中酵母菌在系统启动和稳定后的多样性变化, 结果表明酵母菌在采油废水处理系统中有较高的多样性和稳定性; 原水中具有较高的酵母菌多样性, 并在系统稳定过程中呈现逐渐增加的趋势。表明酵母菌可以在活性污泥系统中稳定存在, 同时酵母菌等真核微生物在烃类污染物去除和环境治理上具有一定的研究和应用 价值。     

酵母菌在疏水性有机污染物去除方面的应用

介绍了酵母菌处理技术在疏水性有机污染物降解中的研究和应用进展,并分析了酵母菌表面特征与其降解活性及在废水处理系统中的稳定性之间的关系。酵母菌技术可以在高含油色拉油废水及含高分于量多环芳烃（PAH）废水的处理中发挥细菌无法替代的作用,酵母菌重要表面特征,特别是其疏水性和乳化能力对于其在疏水性有机污染物处理体系中的稳定性具有重要影响。     

粘红酵母在味精废水中发酵生产油脂

对粘红酵母菌株进行驯化得到一株优良菌株Rh8,利用其在味精废水中发酵以去除废水中的化学需氧量(COD)并生产油脂;考察废水pH以及添加葡萄糖母液、营养因子等对菌株Rh8在味精废水中生长、产油和COD去除效果的影响,发现将废水稀释4倍、调节pH至5.5时,菌株可以较好地生长;而添加废葡萄糖母液、酵母粉、KH2PO4、MgSO4、MnSO4均能够促进菌体的生长、产油和废水中的COD去除,在250 mL摇瓶中,生物量最高可达15.6 g/L,干菌体中油脂质量分数达到29.61%,COD去除率达到45.1%。     

高产γ-氨基丁酸酵母菌株的亚硝基胍诱变选育

目的:探讨亚硝基胍诱变选育高产γ-氨基丁酸酵母菌株的方法。方法:使用亚硝基胍对酵母菌株进行诱变;采用含溴甲酚绿的YEPD培养基筛选突变菌,采用薄层层析法和比色法鉴定变异菌株发酵液中的γ-氨基丁酸及其含量;对突变菌株连续继代培养4代,测定各代发酵液中γ-氨基丁酸的含量,鉴定诱变菌株的遗传稳定性。结果:亚硝基胍诱变酵母的最佳浓度为1.0g.L-1,最佳诱变时间为15min;获得了5株突变菌株,菌落呈绿色;薄层层析法鉴定突变菌株都能产γ-氨基丁酸;诱变菌发酵液中的γ-氨基丁酸含量各异,但高于对照,且增长幅度很大;对突变菌株后代遗传稳定性进行了鉴定,结果表明突变菌株4遗传性较稳定。结论:采用1.0g.L-1的亚硝基胍溶液处理酵母菌15min,经筛选鉴定,获得了一株遗传稳定的高产γ-氨基丁酸的酵母菌株。     

一株分离自酸梅酱的酵母菌的分离鉴定

目的:分离及鉴定来自于新疆塔城民间自制酸梅酱中的酵母菌。方法:用NL1/NL4引物对扩增酵母菌株的26S rDNAD1/D2区,测序结果进行序列分析,用Neighbour-joining(N-J)方法构建系统发育进化树,同时结合酵母的传统形态学鉴定对菌株进行鉴定。结果:26S rDNA D1/D2区序列分析表明菌株KKS与Metschnikowia aff.fructicola D3895相近,相似率为99.2%。在N-J法构建的系统发育进化树中,菌株KKS与Metschnikowia aff.fructicola聚类在同一分枝上。结论:从新疆塔城民间自制酸梅酱中分离得到一株酵母菌并将该菌株鉴定为Metschnikowia aff.fructicola。     

高产Y-氨基丁酸酵母菌株的亚硝基胍诱变选育

目的：探讨亚硝基胍诱变选育高产Y-氨基丁酸酵母菌株的方法。方法：使用亚硝基胍对酵母菌株进行诱变;采用含溴甲酚绿的YEPD培养基筛选突变菌,采用薄层层析法和比色法鉴定变异菌株发酵液中的Y-氨基丁酸及其含量;对突变菌株连续继代培养4代,测定各代发酵液中Y-氨基丁酸的含量,鉴定诱变菌株的遗传稳定性：结果：亚硝基胍诱变酵母的最佳浓度为1．0g．L^-1,最佳诱变时间为15min;获得了5株突变菌株,菌落呈绿色;薄层层析法鉴定突变菌株都能产Y-氨基丁酸;诱变菌发酵液中的Y-氨基丁酸含量各异,但高于对照,且增长幅度很大;对突变菌株后代遗传稳定性进行了鉴定,结果表明突变菌株4遗传性较稳定。结论：采用1．0g．L^-1的亚硝基胍溶液处理酵母菌15min,经筛选鉴定,获得了一株遗传稳定的高产Y-氨基丁酸的酵母菌株。     

海南热带雨林腐木上酵母菌物种多样性研究

用含木糖为唯一碳源(培养基X)和含葡萄糖及7.6%乙醇(培养基E)的两种富集培养基分别从采自海南热带雨林的56和57份腐木样品中分离到酵母菌67和75株.依据26S rDNA D1/D2区域序列分析并结合形态学特征对这些菌株进行了分类学研究,探讨了该地区腐木上的酵母菌物种多样性及其分布.从分离的142株酵母菌中鉴定出14个属63个种,其中疑似新种25个,占总种数的近40%,说明在热带雨林腐木中尚存在大量酵母菌新分类群有待被发现.从用培养基X和E分离的酵母菌中分别鉴定出7属37种和11属33种,优势属均为假丝酵母属Candida Berkhout和毕赤酵母属Pichia Hansen,但种类组成基本不同.用培养基x富集分离的菌株以Candida quercitrusa S.A.Meyer＆Phaff的地理分布最广,用培养基E富集分离的菌株以异常毕赤酵母Pichia anomala(Hansen)Kurtzman、酿酒酵母Saccharomyces cerevisiae Meyen ex Hansen和亚膜毕赤酵母Pichiasubpelliculosa Kurtzman分布最广泛.同一样品用两种不同富集培养基分离的菌株大多数属于不同的种,在对比的23份样品中,只从2份样品中分离到了同一个种的菌株.用培养基X和E分离的菌株分别属于可利用木糖和可耐受乙醇的酵母菌,用两种培养基同时分离到的菌株属于具备利用木糖和耐受高浓度乙醇两种能力的菌株.这些酵母菌在木质纤维素物质的生物乙醇转化技术中的应用价值值得进一步研究.     

耐氟康唑的白假丝酵母菌主动外排机制初探

目的:了解对氟康唑耐药的白假丝酵母菌主动外排系统及主动外排基因CDR1的表达水平。方法:检测氟康唑敏感性和耐药性白假丝酵母菌对罗丹明6G主动外排情况,筛选出主动外排系统功能增强的菌株;采用Northern blot技术检测主动外排系统功能增强的菌株的CDR1基因的表达。结果:在由葡萄糖提供能量的体系中,5株耐药菌株外排罗丹明6G较敏感菌株明显增加,Northern blot发现其中4株CDR1基因表达水平升高。结论:耐氟康唑白假丝酵母菌主动外排基因CDR1表达升高。     

一株假单胞菌HZ-1降解石油烃的初步研究

从石油废水活性污泥中分离到一株可以较好降解石油烃的假单胞菌（Pseudomonas sp）HZ-1,采用生物膜法对该菌株在30℃条件下降解某炼油厂废水中的烃类物质进行了研究.研究结果表明不同营养盐NH4NO3、NaH2PO4、NH4Cl对菌株HZ-1处理石油废水的作用不同,终浓度为1000mg/L的NH4NO3对菌株HZ-1处理石油废水效果最佳.并在30℃和pH 8.0的条件下,初步研究了该菌株降解萘的情况,在萘浓度低于78 mg/L的情况下,120 rpm好氧振荡培养144h,这株菌对萘的降解率在86%以上.     

假丝酵母属疑难菌株大亚基rDNA D1/D2区域序列分析及其分类学意义

对根据常规形态和生理生化性状难以确定分类学地位的8株假丝酵母菌,进行了以大亚基(26S) rDNA中D1/D2区域(约500～600 bp)的碱基序列分析为依据的分子分类学研究.根据系统树上所显示的供试菌株与假丝酵母属及相关子囊菌酵母已知种的亲缘关系,以及与最近缘种模式菌株D1/D2区域序列的相似性比较,确定了各个菌株的归属.本研究也显示了DNA序列分析在假丝酵母菌快速鉴定中的优越性.     

Impacts of cell surface characteristics on population dynamics in a sequencing batch yeast reactor treating vegetable oil-containing wastewater

Ten yeast strains acquired from different sources and capable of utilizing vegetable oil or related compounds (fatty acid or oleic acid) as sole carbon sources were inoculated into a sequencing batch reactor (SBR) for the treatment of high-strength vegetable oil-containing wastewater. The SBR system stably removed >89% of chemical oxygen demand (COD) and >99% of oil when fed with wastewater containing 15 g/L COD and 10 g/L oil in average. Denaturing gradient gel electrophoresis of polymerase chain reaction-amplified 26S rRNA genes showed that among the ten yeast strains, only Candida lipolytica, Candida tropicalis, and Candida halophila were dominant in the system. To elucidate the major factors affecting the selection of yeast strains in the SBR system, the three dominant strains were compared with two non-dominant strains in terms of COD removal performance, biomass yield, cell settleability, cell flocculation ability, cell emulsification ability, and surface hydrophobicity. Results showed that hydrophobicity and emulsification ability of yeast cells were the two most important factors determining the selection of yeast strains in the treatment of high-strength oil-containing wastewater.     

Treatment of distillery wastewater discharged from beet molasses-spirits production using yeast

Treatment of wastewater discharged from beet molasses-spirits production using yeast was studied. Two flocculant strains, Hansenula fabianii J 640-4-1 and Hansenula anomala J 45-N-5, and a strain, I-44, isolated from soil were suitable for treatment of wastewater at the concentration of 47,300 ppm of total organic carbon (TOC). Especially in the case of 2-step treatment using J 45-N-5 and I-44, TOC decreased to 11,900 ppm, and the C/N ratio (TOC/Total nitrogen) decreased from 14.8 to 4.6.     

Characterization of two novel yeast strains used in mediated biosensors for wastewater

After isolation from a pulp mill wastewater treatment facility, two yeast strains, designated SPT1 and SPT2, were characterized and used in the development of mediated biochemical oxygen demand (BOD) biosensors for wastewater. 18S rRNA gene sequence analysis revealed a one nucleotide difference between the sequence of SPT1 and those of Candida sojae and Candida viswanthii. While SPT2 had the highest overall homology to Pichia norvegensis, at only 73.5%, it is clearly an ascomycete, based on BLAST comparisons and phylogenetic analyses. Neighbor-joining dendrograms indicated that SPT1 clustered with several Candida spp., and that SPT2 clustered with Starmera spp., albeit as a very deep branch. Physiological tests, microscopic observations, and fatty acid analysis confirmed that SPT1 and SPT2 are novel yeast strains. Physiological tests also indicated that both strains had potential for use in mediated biosensors for estimation of BOD in wastewater. The lower detection limits of SPT1- and SPT2-based K3Fe(CN)6-mediated biosensors for a pulp-mill effluent were 2 and 1 mg BOD/L, respectively. Biosensor-response times for effluents from eight different pulp mills were in the range of 5 min. Reliability and sensitivity of the SPT1- and SPT2-based biosensors were good, but varied with the wastewater.     

Responses of pea and wheat to textile wastewater reclaimed by suspended sequencing batch bioreactors

Availability of good quality water for crop irrigation is a big challenge in developing countries due to limited resources of clean water. Textile industry consumes a huge amount of water during dyeing process and consequently it releases high strength wastewater into wastewater streams. The present study was designed with the objective to use textile wastewater treated in sequencing batch bioreactor for irrigation purpose. Wastewater containing 100 mg/L reactive black-5 azo dye amended with different co-substrates was treated using mixed liquor suspended solids (MLSS) and two previously isolated dye-degrading bacterial strains (Psychrobacter alimentarius KS23 and Staphylococcus equorum KS26). About 90% color and COD removal in case of dye-containing wastewater amended either with mineral salts + yeast extract or only yeast extract was achieved in 24 h after treatment with mixed culture (MLSS + KS23 + KS26). The treated wastewater was applied for irrigation of pea and wheat plants under controlled conditions. Untreated dye-contaminated wastewater was used as a control for comparison. A significant positive effect of treated dye wastewater amended with different co-substrates on the seed germination index, root and shoot length and biomass was observed in response to application of dye-containing wastewater treated with MLSS and dye-degrading bacterial strains compared to untreated control. Results of this study reveal that the dye-degrading microbial cultures could be used to enhance the treatment efficiency of dye-contaminated wastewater that can be utilized for irrigation of crops and biomass production.     

Yeast strains from the endogenous microflora of the olive fliesBactrocera oleae larvae which could degrade the olive oil mill wastewaters polyphenols

Worldwide, wastewaters constitute a major environmental pollutant. They are very toxic against a wide range of plants and soil microorganisms. Their toxicity is due to the presence of compounds such as polyphenols. In this study, we have isolated yeast strains from the endogenous microflora of the olive fliesBactrocera oleae larvae that were capable of degrading the olive oil mill wastewater polyphenols. The results obtained showed the presence, in the digestive tract of the larvae, of yeast strains resisting to polyphenols. Two resistant strains were isolated and have shown variable capacity of polyphenols degradation that could reach up to 72%. The two isolated strains were identified by two methods: conventional technique and molecular method associating PCR amplification and DNA sequencing of the 5.8S ribosomal RNA gene. Both techniques showed that the two isolated strains corresponded to theCandida diddensiae specie. Related to its capacity to degrade polyphenols, this specie would be a potential candidate for wastewater treatment and environmental protection.     

Anti-infectious potential of beta-mercapto-ethanol treated baker's yeast in gnotobiotic Artemia challenge test

Aim: To evaluate nutritional and anti‐infectious characteristics of the chemically treated baker’s yeast with 2‐mercapto‐ethanol (2ME) for gnotobiotically grown Artemia. Methods and Results: A selection of isogenic yeast strains was treated with 2ME and fed to gnotobiotically grown Artemia. In the first experiment the effect of the chemical treatment on the yeast nutritional value was studied. In most cases, 2ME‐treated yeast cells were better feed for Artemia than the untreated cells. In the second experiment, a small quantity of 2ME‐treated yeast cells was fed to Vibrio campbellii (VC) challenged Artemia. The 2ME‐treatment on some yeast strains (e.g. gas1, kre6 and chs3) significantly improved Artemia resistance against VC compared with the respective untreated yeast cells. Conclusion: Simple chemical treatment with 2ME could significantly improve the nutritional and anti‐infectious properties of some baker’s yeast strains for gnotobiotically grown Artemia. Significance and Impact of the Study: The gnotobiotic Artemia test system provides a unique opportunity (because of noninterference of other microbial compounds) to investigate how the yeast cell wall composition influences macro parameters (e.g. growth and survival) in an organism. In addition, gene expression studies in these gnotobiotically grown Artemia should provide further documentation on direct effects of yeast cells on the genes involved in immune functions.     

Fermentation of starch to ethanol by an amylolytic yeast Saccharomyces diastaticus SM-10

A total of fifteen yeast strains were isolated from natural sources including fruits, soil, molasses, honey and a variety of indigeneous fermented foods. Screening of these strains for growth, ethanol production and glucoamylase activity led to selection of a yeast strain SM-10 identified as S. diastaticus having maximum glucoamylase activity (80 units ml(-1)) and ethanol production from starch (3.5%). Ethanol production from wheat flour was found to be 1.75% which could be increased to 5.2% after treatment of wheat flour with pepsin, diastase and glucoamylase.     

Isolation of the Acrylamide Denitrifying Bacteria from a Wastewater Treatment System Manufactured with Polyacrylonitrile Fiber

Acrylamide has carcinogenicity and toxicity, so its discharge to natural water and soil systems might have an adverse impact on water quality, endangering public health and welfare. The investigation attempts to isolate acrylamide denitrifying bacteria from a wastewater treatment system manufactured with polyacrylonitrile (PAN) fiber. The goal is to elucidate the effectiveness of isolated pure strain and PAN mixed strains in treating acrylamide from synthetic wastewater. The results reveal that Ralstonia eutropha TDM-3 was isolated from the wastewater treatment system manufactured with PAN fiber. The PAN mixed strains and R. eutropha TDM-3 can consume up to 1446 mg/L acrylamide to denitrify from synthetic wastewater. Complete acrylamide removal depended on the supply of sufficient electron acceptors (nitrate). Strain R. eutropha TDM-3, Azoarcus sp. pF6, Azoarcus sp. T, and Herbaspirillum sp. G8A1 are related closely, according to the phylogenetic analyses of 16S rDNA sequences.     

Oxidative stress-resistance assay for screening yeast strains overproducing heterologous proteins

Many natural proteins have been developed into drugs and produced for direct application. Identifying improved hosts to achieve high-level heterologous protein production is a challenge in the study of heterologous protein expression in recombinant yeast. In this study, a novel high-throughput assay to screen such overproducing Saccharomyces cerevisiae strains was systematically developed. The protocol designed was based on screening host strain derivatives with increased superoxide dismutase dependent resistance to oxidative stress. Yeast cells transformed with recombinant plasmid carrying SOD1 gene as a reporter responded exquisitely to oxidative stress induced by elevated concentrations of paraquat. Improved yeast strains resulting from screening clones subjected to genome shuffling through selective pressure argue for a more effective screening system compared with traditonal selection. Moreover, this approach can be employed in general biochemical analysis without utilization of flow cytometry or well plate reader. Therefore, it is expected that the high-throughput assay would make superior strains producing heterologous proteins.     

亚硝化菌株的筛选及其初步鉴定

从化工废水和城镇污水生物处理反应器水中取样,用斯凯尔曼亚硝化假单孢杆菌培养基及硅胶平板分离得到4株可以利用氨氮生长的菌株,并对其形态和生理生化性状进行分析。初步研究结果表明。它们可能分属硝化菌、丝状菌和假单孢菌。在溶氧浓度大于1mg/L的液体培养基中,以上菌株能够将氨氮转化为NO2^-,基本没有NO3^-,又能将氨氮转化为气态氮,且水中NO2^0和NO3^-不再累积,分别保持在小于9mg/L和小于2．0mg/L的水平。