8株窖泥链霉菌对酸、乙醇的适应性研究

对分离自窖泥的8株放线菌进行鉴定,并考察其对窖泥环境中pH值、乙醇的耐受性.采用ISP2液体培养基在pH值2.0～3.5、乙醇浓度3%vol ～6%vol条件下培养链霉菌菌株,结果显示,分属于链霉菌属的8个种的8株放线菌均不能耐受pH值2.0及6%vol的乙醇浓度,均能耐受pH值3.5及3%vol的乙醇浓度,5株链霉菌在pH值3.0时可以生长,2株链霉菌可耐受5%vol乙醇,说明分离自窖泥的多数链霉菌对窖泥环境的适应能力较好,但不同种链霉菌的适应能力存在差异,窖泥环境在长期生产过程中对链霉菌属菌株具有一定的选择驯化作用.     

利用胆固醇氧化酶转化胆固醇制备胆甾-4-烯-3-酮

用酶法催化氧化胆固醇制备胆甾-4-烯-3-酮相对于化学法合成具有反应简单、成本较低等优点.作者探讨了在正辛烷作为有机相的两相反应体系中,以胆固醇氧化酶催化氧化胆固醇制备胆甾-4-烯-3-酮的方法.在胆固醇质量浓度为40g/L,反应时间40min、反应温度40℃、磷酸缓冲液-正辛烷体积比32、通氧40L/h及搅拌转速300r/min下,胆固醇转化率达到92%,经薄板层析及紫外扫描,发现转化产物为单一的胆甾-4-烯-3-酮.     

根霉转化坎利酮的培养基优化

以坎利酮转化率为指标,优化了根霉转化坎利酮的营养需求。通过摇瓶培养和高效液相色谱分析等技术研究了不同碳源、氮源、无机盐以及一些重要因素的浓度对转化率的影响。通过L9(34)正交试验对培养基组合进行优化,优化后的培养条件为:葡萄糖30 g/L,玉米浆25 g/L,酵母膏12 g/L,KH2PO41.5 g/L。在优化后的条件下,坎利酮的转化率为87.68%,较优化前提高9.06%。     

Genome Shuffling Enhanced e-poly-L-lysine Production of a Recombinant Streptomyces sp. Feel-1

利用Genomeshuffling技术对1株融合重组菌Streptomycessp．Feel-1产ε-聚赖氨酸（8-PL）进行了育种研究。首先对Feel-1进行了甲基磺酸乙酯（EMS）诱变,以甘氨酸（Gly）和S-2-氨乙基-L-半胱氨酸（AEC）作为抗性指标进行筛选,得到4株正向突变株,ε—PL摇瓶产量分别为1．78g／L、1．89g／L、1．85g／L、1．86g／L,比原始菌株Feel-1（1．60g／L）分别提高了11．3％、18．1％、15．6％、16．3％;然后将这4株产量提高株作为出发菌库,采用双亲灭活法进行了3轮递推式原生质体融合（Genome shuffling）,最终得到了3株产量大幅提高的重组菌,摇瓶产量分别为2．42g／L、2．35g／L和2．37g／L,比原始菌株分别提高了51．3％、46．9％和48．i％;选择其中1株（G3—67）进行补料分批发酵,192h时ε-PL产量达到32．2g/L。     

Bioconversion of yolk cholesterol by extracellular cholesterol oxidase from Brevibacterium sp.

An efficient process for reducing yolk cholesterol by enzymatic treatment was developed in this paper. Extracellular cholesterol oxidase (COD) from a mutant Brevibacterium sp. ODG-007, showed a strong capacity in bioconversion of yolk cholesterol to cholest-4-en-3-one, especially supplemented with NaCl and lipase C, as a yolk granule solubilizer. The bioconversion process was investigated first, to obtain basic information of the process and was further optimized by analysis of parameters, including COD concentration, dilution ratio and incubation time on the cholesterol conversion, employing Response Surface Methodology (RSM) and Central Composite Design (CCD). Under the optimum operational conditions: COD concentration of 5.39 U/g yolk powder, water: solid ratio of 3.54 and incubation time of 13.75 h, up to 85.6% yolk cholesterol was reduced, and the remaining cholestenone, an effective anti obesity medicine in the product, may raise its commercial value.     

固定化Amycolatopsis sp.ST 2710转化洛伐他汀的研究

研究用海藻酸钠固定化ST2710的技术,确定了较好的制备工艺,并用该固定化细胞对转化洛伐他汀进行了研究。结果表明,在海藻酸钠浓度2%,CaCl2浓度为2%,固化时间为1 h,包埋量为2.5 mL菌悬液/100 mL凝胶的条件下,固定化细胞具有较好的机械强度和较高的转化率。     

链霉菌9-1-1产几丁质酶的发酵条件研究

从烟草根际土样中分离、筛选得到1株几丁质酶活性比较高的链霉菌9-1-1,为评价该菌株利用价值,对其发酵条件(碳源、氮源、初始pH、表面活性剂及发酵时间)进行了研究.结果表明:该菌株的最佳发酵条件以1%胶体几丁质为碳源,发酵液初始pH值为6.5,以1%蛋白胨为氮源,0.1%吐温80作为表面活性剂,发酵时间为96 h,接种量为1%,最高酶活达到56 U/mL.     

链霉菌L2001利用农业废弃物产木聚糖酶条件及酶解产物

本文主要研究产木聚糖醇的菌株对农业废弃物的利用情况并分析酶解产物.对本实验室保藏的木聚糖酶高产菌株进行筛选,其中链霉菌L2001能够较好地利用农业废弃物.通过单因素试验确定最佳碳源、氮源、温度、初始pH等.采用响应面Box-Benhnken中心组合试验设计优化链霉菌L2001产木聚糖酶的条件,同时建立木聚糖酶活力随碳源浓度、pH和温度变化的二次回归方程.结果表明,最佳发酵条件是:棉籽壳3.08%,pH6.48,温度40.1℃.在此条件下,木聚糖酶活力达498.8 U/mL.通过薄层层析分析链霉菌L2001产木聚搪酶的水解产物主要是木二糖和木三糖.     

链霉菌WZFF.L-M1搭载"神舟"四号飞船的空间育种效果

生产微生物谷氨酰胺转胺酶(MTG)链霉菌WZFF.L-M1(酶活2.62U/mL)的孢子和斜面培养菌体搭载“神舟”四号无人飞船,并在斜面中添加诱变剂亚硝基胍(NTG)。实验发现,搭载菌株的菌落形态发生变异,并与产酶能力有一定相关。通过对搭载菌株的初筛与复筛的菌种选育驯化,获得一系列产酶能力大幅度提高的优良突变菌株。其中的3株性能优异,遗传性状稳定,产酶活力提高了30%以上,能够稳定发酵生产高于3.5U/mL的MTG,集中体现了航天飞船搭载的空间诱变育种技术效果。     

Enzymatic Properties of β‐1,3‐Glucanase from Streptomyces sp Mo.

Streptomyces sp Mo endo‐β‐1,3‐glucanase was found to have hydrolyzing activity toward curdlan and released laminarioligosaccharides selectively. The molecular weight was estimated to be 36000 Da and its N‐terminal amino acid sequence was VTPPDISVTN. The optimal pH was 6 and the enzyme was found to be stable from pH 5 to 8. The optimal temperature was 60 °C and the activity was stable below 50 °C. The enzyme hydrolyzed selectively curdlan containing only β‐1,3 linkages. The enzyme had 89% relative activity toward Laminaria digitata laminarin, which contains a small amount of β‐1,6 linkages compared with curdlan, while Eisenia bicyclis laminarin with a higher amount of β‐1,6‐linkages, was not hydrolyzed. Mo enzyme adsorbed completely on curdlan powder. The enzymatic hydrolysis of curdlan powder resulted in the accumulation of laminaribiose (yield 81.7%). Trisaccharide was inevitably released from the hydrolysis of laminarioligosaccharides with 5 to 7 degrees of polymerization (DP). Although the enzyme cleaved off disaccharide (DP 2) from tetrasaccharide (DP 4), the reaction rate was lower than those of DP 5 to 7. The results indicated that the active site of Mo endo‐β‐1,3‐glucanase can efficiently recognize glucosyl residue chain of greater than DP 5 and hydrolyzes the β‐1,3 linkage between the 3rd and 4th glucosyl residue.     

拟无枝酸菌ST2710转化洛伐他汀为无锡他汀的发酵工艺的初步研究

无锡他汀是胆固醇合成途径中限速酶羟甲基戊二酰辅酶A(HMG-CoA)还原酶抑制剂,由洛伐他汀经拟无枝酸菌(Amycolatopsis sp.ST 2710)转化产生。文中在摇瓶水平考察了底物浓度、溶氧和剪切力等因素对洛伐他汀(底物)转化为无锡他汀(产物)的影响,并在5 L发酵罐上进行了分批发酵及材料分批发酵研究。摇瓶结果表明,菌株对溶氧要求较高,对剪切力很敏感,底物适宜添加浓度为1 g/L。在5 L发酵罐间歇发酵过程中。在控制溶氧不低于35%.搅拌转速为300r/min,产物产量达到最高,为0.77 g/L左右,发酵周期为80 h,较摇瓶发酵缩短16 h。采用补料发酵工艺后,有效减缓底物抑制,产物产量达到1.83 g/L。是分批发酵的2.4倍。     

1,3-丙二醇间歇发酵培养基的优化

从Na+、K+、NH4 +等 1价阳离子对甘油发酵生产 1,3-丙二醇过程中关键酶的活性影响分析出发 ,设计改进发酵培养基组成。通过间歇发酵 ,考察甘油初始质量百分比分别为 2 %、4 %和 6 %的发酵情况 ,结果表明 ,采用无铵培养基氨水调节 pH的发酵 ,甘油的转化率和 1,3 丙二醇的终浓度较高 ,而且与文献报道的培养基相比更为简单 ,有利于工业应用。     

产磷脂酶D链霉菌发酵条件优化

以链霉菌为发酵菌株,研究了10 L发酵罐中不同培养条件对链霉菌发酵产生磷脂酶D和链霉菌生长的影响。结果表明,10 L发酵罐中最佳发酵条件为:接种量12%,种龄17 h,温度28℃,通气量0.8 L/h,转速600 r/min。在最佳发酵条件下,酶活可达到3.48 U/m L。     

Kinetics of soybean oil consumption and cephamycin C production in culture of Streptomyces sp. using mineral support

A simple kinetics of soybean oil consumption and cephamycin C production in Streptomyces sp. culture using a mineral support is proposed in this study. The mineral support was used for both suspending the soybean oil as fine oil droplets and immobilizing mycelia. The optimum concentrations of oil and mineral support for obtaining the maximum cephamycin C production were determined to be 50 and 15 g/l, respectively, by the proposed kinetics. At the optimal concentrations, the concentration of cephamycin C estimated from the proposed model and from the experimental data was 2.82 and 2.80 g/l, respectively. The results of the simulation coincided well with the experimental data for various concentrations of the soybean oil and the support. This demonstrates that our model can explain the kinetics of a culture using vegetable oil as the carbon source and mineral support for both oil suspension and mycelial immobilization.     

Streptomyces sp. A22对酱香白酒酿造风味调控的研究

应用从酱香型白酒发酵过程中分离获得的一株产土臭味放线菌（Actinomycetes sp.）A22与产酱香解淀粉芽孢杆菌（Bacillus amyloliquefaciens）B6进行模拟固态发酵,研究其对发酵过程主要特征风味化合物形成的影响及其调控。结果表明,放线菌Streptomyces sp. A22可导致解淀粉芽孢杆菌（B. amyloliquefaciens）B6代谢富集吡嗪类化合物总量降低,对其代谢吡嗪具有调控作用;而B. amyloliquefaciens B6抑制Streptomyces sp. A22代谢富集土臭素,降低发酵糟醅中的异味;Streptomyces sp. A22与B. amyloliquefaciens B6之间存在相互的生物学调节关系,混菌发酵过程可相互抑制其生物量,这是导致其发酵过程风味调控的根本机制,影响酒体中酸类、醇类、酯类等风味物质的形成,改善酒体风味。     

响应曲面法对产蓝色素链霉菌YLA0的高产色素条件优化

以链霉菌YLA0作为出发菌株来优化其高产蓝色素的培养条件,单因素实验的基础上,筛选出C/N比、p H、培养时间三个单因素,利用Box-Behnken的响应曲面设计进一步优化。结果表明,链霉素YLA0高产蓝色素的最佳培养条件为:C/N比为14∶1、p H7.63、培养时间169.69 h,得率11.67 mg/100 m L。该条件下的实际值为优化值的97.04%,说明该模型可靠,可用于链霉菌YLA0高产蓝色素培养条件的优化。      

链霉菌产磷脂酶D发酵培养基的优化

为得到产磷脂酶D的最优培养基组成,以一株分离至土壤的链霉菌为研究对象,以菌体浓度和酶活为测量指标,进行单因素和正交试验。以各种碳源、氮源为考察因素,分析各因素对产磷脂酶D的影响,确定最佳碳源和最佳氮源分别为玉米淀粉、大豆蛋白胨和酵母粉。在单因素试验的基础上进行正交试验,确定最佳碳源和氮源的配比。在正交试验结果基础上,通过单因素实验确定无机盐K2HPO4和MgSO4的添加量。最终确定其最佳配方为:玉米淀粉10 g/L,大豆蛋白胨10 g/L,酵母粉5 g/L,K2HPO42 g/L,MgSO40.3g/L。用该配方培养菌株24 h后,酶活达到最高为3.53 U/mL,是优化前的1.63倍,为产磷脂酶D链霉菌大规模发酵提供了依据。     

重离子诱变对木聚糖酶产生菌产酶的影响

以1株由青藏高原牦牛粪中分离出的链霉菌为出发菌株,该菌株在发酵培养基中能产生胞外木聚糖酶(3 227.346 U/mL)。以此菌株为出发菌株,对其进行重离子辐照诱变处理,从大量突变株中筛选出木聚糖酶高产菌株SZ10-7,其酶活力达到5 338.42 U/mL,与出发菌株相比较,突变株SZ10-7的酶活力提高了1.65倍。对突变株SZ10-7的发酵条件进行了优化研究,结果表明,该菌株的木聚糖酶活力得到进一步提高,达到5 850.20U/mL,其最适发酵条件为:培养基(g/100 mL)为玉米芯∶麸皮(体积比1∶1)5,酵母膏0.8,K2 HPO4.3H2 O 0.1,MgSO4.7H20 0.5,NaCl 0.3,pH 7.0,培养温度25℃振荡培养时间96 h,实验结果表明,重离子辐照诱变技术是一种有效的微生物诱变育种新技术。     

Complete detoxification of tris(1,3-dichloro-2-propyl) phosphate by mixed two bacteria, Sphingobium sp. strain TCM1 and Arthrobacter sp. strain PY1

Tris(1,3-dichloro-2-propyl) phosphate (TDCPP), a flame retardant, is regarded as a potentially toxic and persistent environmental contaminant. We previously isolated a TDCPP-degrading bacterium, Sphingobium sp. strain TCM1, which, however, produced a toxic metabolite: 1,3-dichloro-2-propanol (1,3-DCP). This study was undertaken to develop a technique for complete TDCPP detoxification using strain TCM1 with a 1,3-DCP-degrading bacterium, Arthrobacter sp. strain PY1. For efficient detoxification, we designed a resting cell system and examined the effect of freezing and lyophilization treatments for preparation of their resting cells. Results show that treatments had no marked adverse effect on their activities. The TDCPP dephosphorylation by TCM1 resting cells was optimal at 30°C and pH 8.5. Also, 1,3-DCP dehalogenation by strain PY1 resting cells was optimal at 35°C and pH 9.5. Under those respective conditions, the activities were 2.48 μmol h^− 1·OD₆₆₀^− 1 for TDCPP and 0.95 μmol h^− 1·OD₆₆₀^− 1 for 1,3-DCP. Based on these results, we set the reaction temperature to 30°C and pH to 9.0. Then we examined the detoxification of 50 μM TDCPP using mixed resting cells at a final OD₆₆₀ of 0.05 for strain TCM1 and 0.2 for strain PY1. In these conditions, TDCPP was eliminated after 1 h, but some of the resulting 1,3-DCP remained at a constant level. The increase in strain PY1 cells to a final OD₆₆₀ of 4.0 decreased the TDCPP dephosphorylation rate of strain TCM1 cells but achieved complete detoxification of TDCPP during 12 h of reaction.