鱼腥藻HB1017株化能异养生长的研究

以葡萄糖和蔗糖为碳源,检测了六株（种）鱼腥藻的化能异养生产能力。其中鱼腥藻ＨＢ１０１７株化能异养生长较快,鱼腥藻ＨＢ０株化能异养生长缓慢,其余四种鱼腥藻不能进行化能异养生长。鱼腥藻ＨＢ１０１７株能利用果糖、葡萄糖、蔗糖为底物进行化能异养生长,但生长速率依次递减,差别显著。８磅湿热灭菌的果糖和蔗糖,与过滤灭菌的相比,只能维持低得多的化能异养生长速率。然而,８磅湿热灭菌的葡萄糖能维持比过滤法灭菌的高得     

贵州两处茶园溶磷青霉菌的筛选、鉴定及溶磷能力分析

为维持土壤自然完整性、活化利用土壤中难溶性磷,从贵州名茶产地都匀、贵定茶园土壤中筛选高效溶磷真菌,为制备真菌肥料提供菌种资源。利用溶磷指数(SPI)、形态特征和ITS rDNA序列筛选、鉴定菌株,并采用液体摇床培养实验测定鉴定菌株在以磷酸钙、磷酸铁或磷酸铝为唯一磷源的无机磷液体培养基中的溶磷能力。共筛选到7个高效溶磷菌落,经形态观察分属2种菌株,鉴定为微紫青霉(Penicillium janthinellum)和赭绿青霉(Penicillium ochrochloron)。液体培养基接种、摇床培养15 d,微紫青霉菌在以Ca3(PO4)2、Fe3(PO4)2或AlPO4为唯一磷源的上清液中有效磷含量分别为73.47 mg·L^-1、30.93 mg·L^-1和14.00 mg·L^-1,4℃继续保存至30d后对Fe-P和Al-P的溶解量分别达到72.20 mg·L^-1、32.84 mg·L^-1;赭绿青霉菌培养15d的溶磷量分别为30.72 mg·L^-1、4.14 mg·L^-1和1.51 mg·L^-1,30d对Fe-P和Al-P的溶解量分别达到35.19 mg·L^-1和10.98 mg·L^-1。微紫青霉菌溶解无机磷能力明显优于赭绿青霉菌,有望应用于地区缺磷茶园土壤真菌肥料的制备。     

Use of collagen hydrolysate as a complex nitrogen source for the synthesis of penicillin by Penicillium chrysogenum

Optimal conditions for both biomass formation and penicillin synthesis by a strain of Penicillium chrysogenum were determined when using a collagen-derived nitrogen source. Preliminary investigations were carried out in shaken flask cultures employing a planned experimental program termed the Graeco-Latin square technique (Auden et al., 1967). It was initially determined that up to 30% of a conventional complex nitrogen source such as cottonseed meal could be replaced by the collagen-derived nitrogen source without decreasing the productivity with respect to the penicillin yield. In the pilot scale experiments using a 30 l stirred tank type of bioreactor, higher penicillin yields were obtained when 70% of the conventional complex nitrogen source in the form of cottonseed meal was replaced by the collagen hydrolysate. Furthermore, the maximum rate of penicillin synthesis continued for over a longer period when using collagen hydrolysate as a complex nitrogen source. Penicillin synthesis rates were determined using a linear regression.     

罗伊乳杆菌增殖培养基中碳源氮源的优化

目的 提高罗伊乳杆菌的发酵活菌数,以提高发酵产率,降低生产成本。方法 采用光电比浊法与活菌计数法,通过单因素试验和正交设计方法对罗伊乳杆菌的增殖培养基的碳源和氮源进行优化,并且绘制优化前后的生长曲线。结果 罗伊乳杆菌增殖培养基中最佳的碳源、氮源种类与浓度为葡萄糖2.1%、蔗糖3.0%、牛肉膏1.5%、酵母粉1.4%,最佳的培养时间为10 h。结论 通过优化罗伊乳杆菌的增殖培养基,提高了发酵产率,降低了生产成本。     

碳源和氮源对黑盖木层孔菌菌丝生长的影响

研究了四种碳源(蔗糖、乳糖、葡萄糖、可溶性淀粉)和九种氮源(玉米面、麸皮、马铃薯、大豆粉、酵母粉、蛋白胨、硝酸钾、硝酸铵、尿素)对黑盖木层孔菌菌丝生长的影响。从不同代数的菌丝体中提取多糖,并测定多糖含量、分子量分布范围及单糖组成。结果表明:黑盖木层孔菌菌丝生长的最佳碳源为可溶性淀粉,最佳氮源为玉米面,最优碳氮组合为蔗糖和玉米面的组合。不同代数的菌丝体多糖性状基本稳定。     

微紫青霉CBHⅠ酶纤维素结合结构域在大肠杆菌中的分泌型表达及性质研究

为研究纤维素酶纤维素结合结构域的结构与功能 ,进而深入了解天然纤维素的生物降解机制和提高纤维素酶的生物工艺学价值 ,采用 PCR技术体外扩增了携带微紫青霉外切葡聚糖纤维二糖水解酶 ( CBH ) CBD编码区的 DNA片段 ,将 CBD编码区 DNA片段插入带有 Erwiniacarotovora pe1 B前导肽序列的大肠杆菌质粒 p KK- tac- new上进行了表达 .携带微紫青霉 CBDCBH 编码区的大肠杆菌重组菌株 DH5α( p KK- tac- new- 8)产生有活性的分泌型 CBDCBH 蛋白 .SDS-PAGE检测显示所产生的 CBDCBH 蛋白分子量约 1 0 .8k D.在 IPTG诱导下 ,该菌株所产生的CBDCBH 蛋白含量达 45.2 mg/L,且 90 %以上的 CBD蛋白分泌到培养物上清液中 .结晶纤维素 CF-1 1溶液经 CBDCBH 处理后 ,浊度比对照提高了 1 2 8.9% ,天然棉花纤维结构经 CBDCBH 处理后产生一定程度的非水解性降解作用 ,表明微紫青霉 CBDCBH 具有解聚天然结晶纤维素的作用 .     

Cyanide oxygenase and cyanase activities of Pseudomonas fluorescens NCIMB 11764

Abstract Pseudomonas fluorescens NCIMB 11764 is able to utilise cyanide (both KCN and Ni(CN)₄²⁻) as a nitrogen source for growth. Under such conditions cyanide oxygenase activity is induced. When potassium cyanate (KOCN) is supplied as the sole nitrogen source for growth, cyanase activity is induced. It has been demonstrated that these two enzymic activities are physiologically distinct, and are not co-induced under any of the growth conditions tested.     

微紫青霉CBHI酶的CBD蛋白编码区在大肠杆菌中的亚克隆及表达

对插入质粒pUC18－181上的微紫青霉（Penicilliumjanthinellum）CBHI酶的cDNA基因进行一系列DNA体外操作,包括进行序列定向缺失,最后将两末端修饰为平端后进行连接使质粒环化。用得到的产生序列定向缺失的重组质粒转化大肠杆菌JM109。利用CBD能吸附到结晶纤维素上的特性,从随机选取的24个缺失转化子中筛选到一株含CBD编码区的转化子JM109（pUC18C）,所表达的CBD融合蛋白分子量为21kD．JM109（pUC18C）所产生的LacZ-CBD融合蛋白可通过对纤维素的吸附-解吸附过程一步纯化。其IPTG诱导的pNPC酶活力为零,表明该菌已不再具有CBHI酶活力。     

彩绒革盖菌漆酶产酶条件研究

本文研究了碳源、氮源、愈创木酚、香兰素及培养条件对漆酶分泌的影响;结果表明,淀粉作碳源、干酷素作氮源有利于漆酶的分泌,适宜浓度的愈创木酚和香兰素等对漆酶的产生有一定的作用;pH在3.0－8.0的范围内对漆酶的分泌影响差别不大,培养温度,接种量、通气量对漆酶的分泌有较大影响。     

碳源、氮源对异养单针藻Monoraphidium sp. FXY-10油脂积累和脂肪酸组成的影响

研究了碳源与氮源对单针藻Monoraphidium sp. FXY-10异养培养的影响。以BG-11为基础培养基,通过添加不同类型、浓度梯度碳源和氮源,比较分析微藻生物量、油脂积累以及脂肪酸组成。结果表明,以葡萄糖作碳源,硝酸钠为氮源,微藻细胞积累的油脂是理想的生物柴油制备原料。硝酸钠浓度分别为1.00、3.00和5.00 g/L时,对油脂产量影响不显著(P>0.05)。葡萄糖浓度为10.00 g/L,硝酸钠为氮源油脂产量达到实验最高值0.84 g/L,其油脂脂肪酸组成主要由C16:0和C18:1等短链饱和脂肪酸和单不饱和脂肪酸组成,不饱和度值(DU)为61.98,相对偏低。     

Cloning of exoinulinase gene from Penicillium janthinellum strain B01 and its high-level expression in Pichia pastoris

Aims: The aim of this study is to improve exoinulinase production by expression of a cloned exoinulinase gene inuA1 (GenBank accession no. JF961344 ) from Penicillium janthinellum strain B01 in Pichia pastoris. Methods and Results: A full‐length cDNA of exoinulinase gene (inuA1) was cloned from P. janthinellum strain B01 using RACE PCR. An open reading frame (ORF) of 2115 bp is interrupted by a single intron of 67 bp. The fragment encodes a signal peptide with 20 amino acids and a mature protein with 684 amino acids. The inuA1 was subcloned to the pPICZαC expression vector and succesfully over‐expressed in Pichia pastoris X‐33. The highest activity of exoinlinase reached 272·8 U ml^?1 in the fermentation liquid. It was c. 11‐fold of that produced by wild‐strain B01. A large amount of fructose was identified after the hydrolysis of inulin with the crude recombinant exoinulinase. The recombinant exoinulinase was purified and characterized. The molecular weight of the purified recombinant exoinulianse was 100 kDa. The mass spectrometry result indicated that the purified protein was indeed recombinant exoinulinase. The optimal pH and temperature of the purified recombinant exoinulianse were 4·5 and 50°C, respectively. Conclusions: An exoinulinase gene of P. janthinellum strain B01 was cloned, sequenced and over‐expressed successfully in P. pastoris. Significance and Impact of the Study: Only a few genes have been cloned from P. janthinellum because its molecular biology is poorly understood. In this study, we cloned and over‐expressed inuA1 gene of P. janthinellum in P. pastoris. This recombinant exoinulinase can be used to hydrolyse inulin to produce fructose and facilitate the biofuel production from inulin resources.