采用响应面法优化木糖醇发酵培养基

将Plackett-Burman和响应面设计相结合,对木糖醇发酵培养基进行了优化。结果表明,初始木糖浓度、酵母膏添加量以及MgSO4.7H2O浓度是影响木糖醇转化率的主要因素。优化得到的培养基组成为(g/L)木糖100.7,酵母膏5.302,NaCl6.0,MgSO4.7H2O0.379,KH2PO43,(NH4)2HPO44;通气条件为装液量100mL/250mL。此条件下木糖醇的转化率为0.784g/g。     

响应面法优化不同型态热带假丝酵母发酵生产木糖醇的工艺

利用Design Expert软件对菌丝型和酵母型热带假丝酵母发酵生产木糖醇实验进行设计及结果分析,建立木糖和木糖醇浓度与4个关键因子(菌型、发酵温度、pH、初始木糖浓度)的二次多项式回归模型,并对模型进行解析。结果表明:菌丝型热带假丝酵母转化木糖为木糖醇的能力高于酵母型;升高发酵温度,有利于木糖转化为木糖醇,而pH升高对转化过程并没有明显促进;发酵液中初始木糖浓度与木糖转化率呈正相关关系;获得最佳发酵工艺条件为菌种采用菌丝型酵母,发酵温度37℃,pH8,初始木糖浓度60mg/mL,此时木糖醇浓度达到17.21mg/mL。     

Assignment of most genes encoding major peroxisomal polypeptides to chromosomal band V of the asporogenic yeast Candida tropicalis

The peroxisomes of the asporogenic yeast Candida tropicalis contain about 20 major polypeptides (PXPs). We have isolated a number of genes encoding them; 11 POX genes encoded independent PXPs and three POY genes were likely to encode three other PXPs. To locate these genes on the chromosomes, chromosomes of C. tropicalis were separated by pulsed-field gel electrophoresis. Eight chromosomal bands were observed over the range of 1.0 Mbp (band 1) to 2.8 Mbp (band VIII); the genome size was estimated to be about 20 Mbp. Southern blot analysis showed that ten genes were on band V, three genes were on band IV, and the other gene was on band VI. Three genes gave hybridization signals of nearly equal intensity on two different chromosomal bands: POX6A and POX8B, on bands V and VII; and POX8A, on bands IV and VI. Ribosomal RNA genes also hybridized to two bands, VI and VII. Most genes assigned to only one band hybridized to two restriction fragments produced by either NotI or SfiI endonuclease. The results suggested that C. tropicalis was diploid and that restriction sites were conserved little between homologues. The three POX genes that were found on two chromosomal bands hybridized to not more than two restriction fragments, implying that the allelic genes were present on different chromosomal bands.     

富核糖核酸热带假丝酵母发酵培养基优化

以热带假丝酵母（Candida tropicalis）AY91009为试验菌株,以其核糖核酸（RNA）含量为目标,选取糖蜜为碳源,采用响应面法对其发酵培养基进行优化,建立酵母浸粉、NH4Cl和ZnSO4·7H2O的二次回归模型,确定培养基最佳配方为：糖蜜（30%含糖量）140 mL/L、酵母浸粉2.90%、NH4Cl 1.37%、NaH2PO4·2H2O 0.10%、MgSO4·7H2O 0.20%、FeSO4·7H2O 0.05%、ZnSO4·7H2O 0.10%。在此优化培养基中发酵培养12 h,RNA含量达到11.58%,比优化前提高了35.6%。20 L罐分批发酵试验结果表明,细胞干质量达到32.38 g/L,RNA含量为6.69%,总RNA含量为2.17 g/L,为后续的高密度发酵研究奠定了良好的基础。     

Lipid composition and cell surface hydrophobicity of Candida albicans influence the efficacy of fluconazole–gentamicin treatment

Adherence of the fungus, Candida albicans, to biotic (e.g. human tissues) and abiotic (e.g. catheters) surfaces can lead to emergence of opportunistic infections in humans. The process of adhesion and further biofilm development depends, in part, on cell surface hydrophobicity (CSH). In this study, we compared the resistance of C. albicans strains with different CSH to the most commonly prescribed antifungal drug, fluconazole, and the newly described synergistic combination, fluconazole and gentamicin. The hydrophobic strain was more resistant to fluconazole due to, among others, overexpression of the ERG11 gene encoding the fluconazole target protein (CYP51A1, Erg11p), which leads to overproduction of ergosterol in this strain. Additionally, the hydrophobic strain displayed high efflux activity of the multidrug resistance Cdr1 pump due to high expression of the CDR1 gene. On the other hand, the hydrophobic C. albicans strain was more susceptible to fluconazole–gentamicin combination because of its different effect on lipid content in the two strains. The combination resulted in ergosterol depletion with subsequent Cdr1p mislocalization and loss of activity in the hydrophobic strain. We propose that C. albicans strains with different CSH may possess altered lipid metabolism and consequently may differ in their response to treatment.     

发酵木糖生产乙醇菌株的筛选与鉴定

从花圃土和腐朽树皮中各筛选得到一株可以发酵木糖产生乙醇的菌株。乙醇产量分别可达到6.98g/L和6.05g/L。经过18SrDNA同源性的分析,菌株3鉴定为间型假丝酵母（Candida intermedia）,而菌株621则为热带假丝酵母（Candida tropicalis）。对其发酵木糖、葡萄糖和混合糖的产物蒸馏后用气相色谱分析,主产物为乙醇,纯度高,副产物少。两菌株为下一步的菌种基因改造工作提供了研究基础。     

热带假丝酵母发酵法生产木糖醇的研究

目的利用热带假丝酵母研究发酵木糖生产木糖醇的发酵条件。方法采用摇瓶发酵对发酵生产条件,如培养基中初始木糖浓度、接种量和通气量、氮源、pH等进行优化,通过测定发酵液中木糖的残留量、木糖醇的转化率来确定适合的发酵工艺。结果通过实验得到最佳培养基条件为:初始木糖50g/L,蛋白胨5g/L,酵母粉10g/L,硫酸镁0.5g/L、磷酸二氢钾5g/L,硫酸铵1g/L;最佳发酵条件为:pH 6.0,摇瓶发酵装液量50mL/250mL,转速200 r/min,发酵温度30℃,发酵时间28h。结论优化了木糖醇的发酵工艺。     

一株热带假丝酵母木糖醇高产新菌株的筛选

自木糖厂环境样品中分离出一株编号为1-18的产木糖醇分离物。在初始木糖浓度200g/L的条件下重复循环利用1-18细胞摇瓶发酵,木糖醇转化率高达0.91(木糖醇g/木糖g),接近理论极限值;木糖醇生成速率达6.7g/L·h。无论产物浓度,产物生成速率与产物转化率都达到了令人满意的高水平。1-18发酵液以40ml/kg·d的最大剂量饲喂小白鼠一周,受试动物未出现中毒反应。分离物1-18在木糖醇生产中显然具有重要的生物工程应用潜力。根据常规形态鉴别,生理生化实验测定,以及26S rDNA D1/D2区域核酸序列同源性比较分析,证实分离物1-18属于Candida tropicalis(热带假丝酵母)。分离物1-18已保存于中国典型培养物保藏中心,保藏编号CCTCC M 205067,其26S rDNA D1/D2区域核酸序列在GenBank的核酸序列登记号为DQ176428。     

热带假丝酵母木糖醇脱氢酶的分离纯化

探讨了热带假丝酵母中木糖醇脱氢酶的分离纯化工艺。分步采用硫酸铵分级沉淀、DEAE-Sepharose离子交换层析、Sephacryl S-200凝胶层析以及Hi Trap Blue HP亲和层析等纯化方法,得到电泳纯的木糖醇脱氢酶。经SDS-PAGE和native-PAGE分析表明,木糖醇脱氢酶分子质量约为90ku,由2个分子质量为45ku的亚基组成。最终酶蛋白纯化倍数达到60·8倍,回收率为10·9%。     

响应面法优化酒糟水解液发酵生产木糖醇

研究热带假丝酵母(Candida tropicalis)GIM2.7发酵酒糟水解液生产木糖醇的最佳工艺条件,提高酒糟资源的利用率。在单因素试验结果的基础上,选择主要因素接种量、装液量、有机氮源添加量进行响应面优化。结果表明,当接种量为16%、有机氮源添加量5%、装液量109 mL/250 mL时,木糖醇转化率可达到57.8%,与预测值(60.4%)相接近,说明响应面优化酒糟水解液生产木糖醇的发酵条件是有效的。     

热带假丝酵母发酵生产木糖醇的研究

对热带假丝酵母 (C tropicalis )As2 1 776发酵木糖醇的营养条件进行了初步研究。初始木糖浓度在 80g/L附近时木糖醇转化率较高 ,限制性供氧条件下有利于木糖醇积累。酵母膏和蛋白胨是比较适合产木糖醇的有机氮源 ,而酵母膏更利于酵母细胞生长。培养基中添加 2 g/L的(NH4 ) 2 HPO4 、2～ 6g/L的NaCl、1～ 3g/L的KH2 PO4 、0 1～ 0 3 g/L的MgSO4 ·7H2 O能提高木糖醇的转化率     

双歧杆菌在不同材料上形成生物膜的研究

目的:评价双歧杆菌生物膜生成能力.方法:采用结晶紫染色和刚果红染色等方法对几种不同双歧杆菌形成生物膜的能力进行测定和鉴定.结果:采用不同途径分离的9株双歧杆菌分别在亲水性材料和疏水性材料上展现不同的生物膜形成能力.所有菌株在聚丙烯材料上均表现出弱的生物膜形成能力,而在聚苯乙烯和硼硅酸盐材料表面均表现出不同程度的生物膜形成能力.利用盖玻片对生物膜的显微观察表明生物膜含有多糖成分.细胞表面疏水性研究表明双歧杆菌中生物膜的形成与细胞表面疏水性未见相关性.结论:双歧杆菌具有一定的生物膜生成能力.     

Cell surface hydrophobicity and colony morphology of Trichosporon asahii clinical isolates

Trichosporon asahii is a pathogenic basidiomycetous yeast. Individual strains of T. asahii have different colony morphologies. However, it is not clear whether cell surface phenotypes differ among the colony morphologies. Here we characterized the cell surface hydrophobicity and analysed the carbohydrate contents of the cell surface polysaccharides in T. asahii clinical isolates with various colony morphologies. Among the three distinctive colony morphologies obtained from one clinical isolate, the white‐type morphology exhibited higher hydrophobicity. The hydrophobicity of heat‐killed T. asahii cells was greatly reduced after periodate oxidation of the cell surface carbohydrates. Furthermore, the cell wall and extracellular polysaccharide components differed among the morphologies. Our results suggest that T. asahii cell surface hydrophobicity is affected by cell surface carbohydrate composition. Copyright © 2016 John Wiley & Sons, Ltd.     

大豆糖蜜高产单细胞蛋白菌株的筛选及其生长条件

以大豆糖蜜为底物,通过平板初筛和三角瓶摇瓶发酵复筛,从8株单细胞蛋白生产常用的酵母菌中筛选出在10%大豆糖蜜中生物量和总蛋白产量最高的热带假丝酵母CGMCC2.587,其细胞生物量为8.4125g/L,总蛋白产量为4.4696g/L。通过单因素实验确定了菌株生长的最佳培养基为5%麦芽汁培养基;培养条件为初始pH5.0、装液量50mL/250mL、转速160r/min、温度30℃。     

热带假丝酵母利用酒糟水解液发酵生产木糖醇的初步研究

对热带假丝酵母(C.tropicalis)AY91009利用酒糟(丢糟)水解液发酵木糖醇进行了初步研究。结果表明:最佳发酵时间48h,最佳种子龄22h。摇瓶分批发酵工艺条件的最佳组合是:起始pH5.5,接种量15%(v/v),装液量135mL,氮源加入量为10mL含有10g/L酵母膏和20g/L蛋白胨的有机氮源。除水解液本身含有的木糖外,1/10的葡萄糖加入量(w/w)有利于菌体生长和木糖醇的转化,蔗糖则会抑制木糖醇的生成。培养基中添加6g/L的NaCl、3g/L的KH2PO4、0.2g/L的MgSO4.7H2O有利于木糖醇的积累。     

十二碳二元酸产生菌热带假丝酵母的诱变

以热带假丝酵母菌(Candida tropicalis)为出发菌株,经紫外线和氯化锂复合诱变或亚硝酸诱变获得突变株。通过摇瓶分批发酵,对菌株诱变前后发酵产长链二元酸量进行了测定。筛选出能够发酵产酸的突变株12株,其中8株表现出正突变,菌株最高产酸量提高了9.16%。     

热带假丝酵母转化生产木糖醇条件优化

利用热带假丝酵母(Candida tropicalis)生物转化木糖醇工艺简单、能耗低并且产物稳定.该文研究了菌体培养时间、转化时间、pH值、初糖浓度、摇床转速、加入菌体量对转化率的的影响,从而确立了最佳条件,即菌体培养时间为16h、转化时间为10h、转化pH值为5.5、转化初糖浓度为20g/L、转化摇床转速为150r/min、加入菌体量为10%(v/v),优化后木糖醇生物转化率达90%.