促进剂对发酵生产辅酶Q10的影响

研究了几种促进剂对放射型根瘤菌细胞生长及其辅酶Q10发酵的影响。结果表明:在培养基中适量添加VB1、乙酸钠、花生油均可明显提高辅酶Q10的产量,使用正交试验优化后的促进剂组合可使辅酶Q10产量达到50.7 mg/L,比对照组提高33%。     

利用Pichia pastoris生产S-腺苷甲硫氨酸的发酵工艺

在摇瓶中考察了重组Pichia pastoris发酵的诱导剂量,L-甲硫氨酸,以及pH对腺苷甲硫氨酸产量的影响.放大到3.7 L发酵罐和30 L发酵罐后,研究了重组细胞的发酵过程变化,对S-腺苷甲硫氨酸初步纯化.摇瓶中优化后的发酵条件是:每天添加1%甲醇诱导,L-甲硫氨酸为50mmol/L,培养基pH 5.0.培养144 h后SAM产量达到2.32 g/L.3.7 L发酵罐中发酵251 h后细胞浓度为120 g/L,SAM总量为15.18 g.放大到30 L发酵罐中,发酵225.5 h后细胞浓度约为120 g/L,SAM总量为145.05 g.纯化后SAM的纯度为93.5%,回收率为84.5%.     

辅酶Q_(10)产生菌发酵条件的优化

对辅酶Q10生产菌株鞘氨醇单胞菌YZ0803的发酵条件进行优化,确定发酵时间为90 h,250 mL摇瓶装液量为30 mL。培养基组成(质量分数,下同):葡萄糖1.5%,淀粉2.5%,黄豆饼粉2.5%,(NH4)2SO40.5%,NaCl0.03%,K2HPO40.02%,MgSO40.005%。优化后的辅酶Q10产量达到192 mg/L,比采用基础培养基的产量(138mg/L)提高了39.13%。     

一种增加毕赤酵母生产胰岛素前体的方法

为了提高胰岛素前体(PI)的产量,构建了p PIC9K-PI表达载体并电转化至毕赤酵母菌株GS115中,在浓度为4.0 mg/m L的G418抗性平板上筛选到了1株拷贝数为12的菌株CL012。将SNAREs(可溶性N-乙基马来酰亚胺敏感因子受体蛋白)组分中的SNC2和SNC2-SSO2分别转入菌株CL012中,并在摇瓶和5 L发酵罐水平上检测SNAREs对PI产量的影响。结果表明:摇瓶水平上,甲醇诱导96 h后,菌株CL012的PI产量为1.53 mg/L;表达SNC2和SNC2-SSO2的菌株的PI产量分别为1.89 mg/L和2.21 mg/L,分别比菌株CL012提高了23.53%和44.44%。在5 L发酵罐上进行高密度发酵,甲醇诱导96 h后菌株CL012的PI产量为53 mg/L,是摇瓶水平的34.64倍;表达SNC2和SNC2-SSO2的菌株的PI产量分别达到64 mg/L和78 mg/L,分别比菌株CL012提高了20.75%和47.17%。由此得出结论 SNAREs可以促进胰岛素前体的分泌,从而提高在毕赤酵母中的异源表达。     

海洋中海藻糖产生菌的筛选及发酵条件优化

从180余份海水、海泥样品中筛选得到60株产海藻糖较高的菌株,编号为2-14的菌株海藻糖产量最高,为127.9mg/g cell。对2-14菌株进行形态特征、培养特征及生理生化试验,鉴定该菌株为红酵母属(Rhodotorula sp.)。研究摇瓶发酵条件对红酵母海藻糖产量的影响,结果为:初始pH5.5,发酵温度28℃,装液量75mL(250mL三角瓶中)。采用优化后发酵条件红酵母海藻糖产量为193.3mg/g cell,优化前对照值为132.1mg/g cell,优化后的结果是优化前的1.46倍。在5L发酵罐中培养得到最佳发酵时间为54h,发酵罐培养发酵液中海藻糖含量最高达2.5g/L,为摇瓶培养的1.6倍。     

发酵生产辅酶Q10高产菌株的选育

以实验室保存的类球红细菌(Rhodobacter sphaeroides)JDW61为出发菌株,考察了紫外、紫外结合氯化锂和亚硝基胍对菌株产生辅酶Q10能力的诱变效应,并结合辅酶Q10的合成途径设计了快速筛选辅酶Q10高产菌株的模型,获得一株辅酶Q10产量提高的突变株CP222,该菌株摇瓶发酵的辅酶Q10产量为276.14mg·L-1,较出发菌株提高了190%,并且遗传性能稳定。     

培养方式对富硒产朊假丝酵母性能的影响

在摇瓶和5 L发酵罐水平上分别考察亚硒酸钠浓度及其添加方式对高性能(高有机硒含量和高谷胱甘肽含量)富硒产朊假丝酵母制备的影响.结果表明:亚硒酸钠添加质量浓度为15 mg/L时,产朊假丝酵母具有较好的富硒效果,但一次性添加对酵母细胞有较大的毒害作用.采用分批次添加亚硒酸钠的方法获得了较好的制备高性能富硒产朊假丝酵母的培养方式:发酵起始添加L-蛋氨酸10 mmol/L,并在发酵过程的12和15 h分别添加亚硒酸钠10和5 mg/L.在此培养方式下,产朊假丝酵母胞内谷胱甘肽和有机硒含量分别达到172.3 mg/L和1194 μg/g.     

乳酸钠促进法夫酵母虾青素合成代谢流作用分析

【目的】研究乳酸钠(一种糖代谢产物)的加入对法夫酵母JMU-VDL668发酵过程中细胞生长和虾青素合成的影响。【方法】分别在摇瓶和7 L发酵罐实验基础上,采用代谢通量分析的方法分析添加乳酸钠对法夫酵母菌株JMU-VDL668合成虾青素代谢流的影响。【结果】在7 L发酵罐实验中添加乳酸钠,虾青素产量最高可达17.70 mg/L,与对照组相比提高26%。代谢通量分析表明,乳酸钠可以调节丙酮酸、乙酰辅酶A节点处的代谢通量分布,乳酸在乳酸脱氢酶的作用下可以直接进入代谢网络的后半程,乙酰辅酶A的通量和进入TCA循环的通量得到了显著加强。【结论】乳酸钠的加入提供了更多的乙酰辅酶A等前体物质和能量供给,因此促进了虾青素的合成。     

茄尼醇转化高产辅酶Q10菌株的分离鉴定与发酵条件的研究

采用以异戊二烯为唯一碳源的选择性平板筛选模型,从钱塘江沿岸杭州市九堡段土壤中新筛选到一株产辅酶Q10的细菌菌株E03,经形态、生理生化、Biolog碳源利用试验和16S rDNA序列分析,确定E03属于鞘氨醇属(Sphingomonas sp.),命名为Sphingomonas sp.ZUTE03.摇瓶试验确定了该菌发酵生产辅酶Q10的最佳碳源为葡萄糖15 g/L,氮源为硫酸铵10 g/L,初始pH8.0,发酵温度25℃,并考察了该菌转化茄尼醇产辅酶Q.0的发酵工艺,以合适溶剂为溶解体系,于发酵培养基中摇床培养12 h后,加入终浓度为0.75 g/L的茄尼醇粗品,转化12 h,辅酶Q10产值可达96.88 mg/L.     

毕赤酵母Gpn12异戊烯基转移酶NovQ催化合成MK-3

对异戊烯基转移酶NovQ在毕赤酵母Gpn12异源表达过程中诱导剂甲醇添加量进行了探究,并以毕赤酵母Gpn12全细胞为酶源,以甲萘醌、异戊烯醇为前体,催化合成维生素K2(MK-3)。每24 h添加2%甲醇时,NovQ表达量提高约36%。考察摇瓶中初始pH、温度、甲醇添加量、前体(甲萘醌、异戊烯醇)添加量、催化时间、十六烷基三甲基溴化铵(CTAB)添加量等7个因素对Gpn12全细胞催化合成MK-3的影响,发现催化温度、甲萘醌添加量、催化时间影响显著,对3个显著因素进行响应面优化得出催化条件为:催化温度31.56℃,甲萘醌添加量295.54 mg/L,催化时间15.87 h,优化后的摇瓶MK-3产量达到98.47 mg/L,与响应面预测结果一致,较优化前对照组提高了35%。在30 L发酵罐进行生物催化实验,催化时间24 h,细胞催化剂浓度220 g(干重)/L,MK-3产量达到189.67 mg/L。该方法为Gpn12规模化生产MK-3奠定了一定的基础。     

Association of colony morphology with coenzyme Q(10) production and its enhancement from Rhizobium radiobacter T6102W by addition of isopentenyl alcohol as a precursor

Rhizobium radiobacter T6102 was morphologically purified by the aniline blue agar plates to give two distinct colonies; white smooth mucoid colony (T6102W) and blue rough colony (T6102B). The coenzyme Q(10) (CoQ(10)) was produced just by T6102W, showing 2.0 mg/g of CoQ(10) content, whereas the T6102B did not produce the CoQ(10). All of the used CoQ(10) biosynthetic precursors enhanced the CoQ(10) production by T6102W. Specifically, the supplementation of 0.75 mM isopentenyl alcohol improved the CoQ(10) concentration (19.9 mg/l) and content (2.4 mg/g) by 42% and 40%, respectively.     

Coenzyme Q10 production by Sphingomonas sp. ZUTE03 with novel precursors isolated from tobacco waste in a two-phase conversion system

Coenzyme Q10 (CoQ10) is a widely used supplement in heart diseases treatment or antioxidative dietary. The microbial production of CoQ10 was enhanced by addition of solanesol and novel precursors recovered from waste tobacco. The novel precursors were separated by silica gel and identified as alpha-linolenic acid (LNA) and butylated hydroxytoluene (BHT) based on the effect on CoQ10 production and GC-MS. The effects of novel precursors on CoQ10 production by Sphingomonas sp. ZUTE03 were further evaluated in a two-phase conversion system. The precursor's combination of solanesol (70 mg/l) with BHT (30 mg/l) showed the best effect on the improvement of CoQ10 yield. A maximal CoQ10 productivity (9.5 mg l-1 h-1) was achieved after 8 h conversion, with a molar conversion rate of 92.6% and 92.4% on BHT and solanesol, respectively. The novel precursors, BHT and LNA in crude extracts from waste tobacco leaves, might become potential candidates for application in the industrial production of CoQ10 by microbes.     

营养条件和流加发酵对放射型根瘤菌(Rhizobium radiobacter)产辅酶Q10的影响

利用放射型根瘤菌WSH2 6 0 1(RhizobiumradiobacterWSH2 6 0 1)重点考察了葡萄糖、蔗糖、玉米浆和蛋白胨、添加物以及流加发酵对细胞生长和产辅酶Q1 0 的影响 ,结果表明 ,葡萄糖和蔗糖适合于生产辅酶Q1 0 的最佳浓度分别为 30g L和 40g L ;辅酶Q1 0 发酵时玉米浆和蛋白胨的最适浓度分别为 11g L和 16g L ;添加蕃茄汁、玉米浆能提高发酵液的生物量 ,玉米浆、异戊醇、L 甲硫氨基酸等能促进辅酶Q1 0 的积累 ;与分批发酵相比 ,在 7L罐上流加蔗糖其细胞生物量 (DCW)和辅酶Q1 0 积累量增加 ,若在流加蔗糖的同时流加适当浓度的玉米浆能显著提高辅酶Q1 0 的产量 ,最大产量达到 5 2 .4mg L ;最大生物量 (DCW)和胞内辅酶Q1 0 含量 (C B值 )分别达到 2 6 .4g L和 2 .38mg g DCW ,比不流加的分批发酵分别提高 5 3 %和 33% ,比只流加蔗糖分别提高 2 4%和 2 6 %。     

Coenzyme Q(10) production by immobilized Sphingomonas sp. ZUTE03 via a conversion-extraction coupled process in a three-phase fluidized bed reactor

A three-phase fluidized bed reactor (TPFBR) was designed to evaluate the potential of CoQ(10) production by gel-entrapped Sphingomonas sp. ZUTE03 via a conversion-extract coupled process. In the reactor, the CoQ(10) yield reached 46.99 mg/L after 8 h of conversion; a high-level yield of about 45 mg/L was maintained even after 15 repetitions (8 h/batch). To fully utilize the residual precursor (para-hydroxybenzoic acid, PHB) in the aqueous phase, the organic phase was replaced with new solution containing 70 mg/L solanesol for each 8 h batch. The CoQ(10) yield of each batch was maintained at a level of about 43 mg/L until the PHB ran out. When solid solanesol was fed to the organic phase for every 8 h batch, CoQ(10) could accumulate and reach a yield of 171.52 mg/L. When solid solanesol and PHB were fed to the conversion system after every 8 h batch, the CoQ(10) yield reached 441.65 mg/L in the organic phase after 20 repetitions, suggesting that the conversion-extract coupled process could enhance CoQ(10) production in the TPFBR.     

埃博霉素B小试发酵罐发酵条件研究

研究了纤维堆囊菌(Sorangium cellulosum)So F5-76在5 L发酵罐水平上发酵生产埃博霉素B的基本工艺参数,具体考察了接种量、搅拌转速、通气量、添加消泡剂及补糖等5个工艺参数对埃博霉素B发酵产量的影响。最后确定发酵罐基本发酵条件为接种量9%,搅拌转速180 r/min,空气流量3.5 L/min,消泡剂种类选择Antifoam B聚醚类消泡剂,补糖控制在发酵液糖浓度为0.2 g/L,在此条件下埃博霉素B的产量可达25.6 mg/L。     

对沼泽红假单胞菌生产CoQ10添加前体物的优化

CoQ10因其具有多种生理生化功能而不仅用于药物也用作食品添加剂.本研究采用全因析中心复合设计及响应面法对茄尼醇、对羟基苯甲酸及甲硫氨酸三种CoQ10前体物的添加量进行了优化,以达到沼泽红假单胞菌J001最大量地生产CoQ10的目的.结果表明:经过对所得模型进行响应曲面法分析,当茄尼醇、对羟基苯甲酸及甲硫氨酸最佳添加量分别为124.8mg1-1,267.7mg1-1,130.2mg1-1时,得最大CoQ10产量预测值40.6[(mgCoQ10)(g干细胞)-1].对上述最佳组合进行试验验证得40.5±0.2[(mg CoQ10)(g干细胞)-1],很接近预测值,比对照(未加三种CoQ10前体物)CoQ10产量提高了109.8%.     

Effects of dietary L-carnitine and coenzyme Q10 supplementation on performance and ascites mortality of broilers

The study was conducted to determine the effects of dietary L-carnitine and coenzyme Q10 (CoQ10) supplementation on growth performance and ascites mortality of broilers. A 3 x 3 factorial arrangement was employed with three levels (0, 75 and 150 mg/kg) of L-carnitine and three levels of CoQ10 (0, 20 and 40 mg/kg) supplementation during the experiment. Five hundred and forty one-day-old Arbor Acre male broiler chicks were randomly allocated into nine groups with six replicates each. All birds were fed with the basal diets from day 1 to 7 and changed to the experimental diets from day 8. During day 15 to 21 all the birds were exposed to low ambient temperature (15-18 degrees C) to induce ascites. The results showed that under this condition, growth performance of broilers were not significantly affected by CoQ10 or L-carnitine + CoQ10 supplementation during week 0-3 and 0-6, but body weight gain (BWG) of broilers was significantly reduced by 150 mg/ kg L-carnitine during week 0-6. Packed cell volume (PCV) of broilers was significantly decreased by L-carnitine and L-carnitine + CoQ10 supplementation (P < 0.05). Erythrocyte osmotic fragility (EOF), ascites heart index (AHI) and ascites mortality of broilers were significantly decreased by L-carnitine, CoQ10 and L-carnitine + CoQ10 supplementation. Though no significant changes were observed in total antioxidative capability (T-AOC), total superoxide dismutase (T-SOD) was increased by L-carnitine, CoQ10 and L-carnitine + CoQ10 supplementation (P < 0.05). Malonaldehyde (MDA) content was significantly decreased by CoQ10 and L-carnitine + CoQ10 supplementation. The results indicate that dietary L-carnitine and CoQ10 supplementation reduce ascites mortality of broilers; the reason may be partially associated with their antioxidative effects.     

辅酶Q_(10)的毒性试验及对溶血空斑形成细胞的刺激效应

辅酶Q_10（以下简称CoQ_(10)）是生物体内普遍存在的一类生物活性物质,具有重要的生理、生化作用。它是细胞呼吸链中主要的递氢体,在生物能量转化中占重要地位,它又能作为非特异性免疫增强剂,增强机体的防御能力,因此人们对于CoQ_(10)的临床研究越来越受到重视（中国科学院昆明动物研究所,1979）。 我们结合我国具体情况,独创地从提取细胞色素丙的猪心残渣中分离制备了CoQ_(10), （许以盛等,1985）,并与有关生产及临床单位协作,于1977年12月通过了鉴定,为我国的生化药物填补了一项空白。     

氩离子激光照射对类球红细菌的诱变效应及对辅酶Q10产生量的影响

以低产量辅酶Q10类球红细菌为亲本,以氩离子激光为诱变源,对其幅照诱变,结果发现:亲本株发生了明显的诱变效应,出现了不同的色素突变表型。诱变后的色素突变株不仅遗传性状稳定,且辅酶Q10产量比亲本株有明显提高。对其中的黄色突变株发酵液进行辅酶Q10提取及测定,结果显示:其辅酶Q10产量比亲本株提高102.10%,经发酵条件初步优化,其最高产量可达26.39 mg/L发酵液。