灰黄霉素菌的诱变效应

以灰黄霉素产生菌D－756为出发菌株,经过三代紫外线＋氯化锂诱变处理,获得耐氯变株F－1012,该变株在形态特征及产量、耐氯性等方面均发生变化,当发酵培养基中的氯化物浓度由1.5%提高到2.0%,F－1012的大米孢子效价提高了34.5%;当固体平板培养基中氯化物浓度提高到11％时,F－1012的孢子存活率比出发菌株提高了25.5％。     

钾细菌诱变选育和发酵的研究

通过钾细菌的诱变选育和发酵工艺探索 ,来提高发酵菌量和芽孢量 ,确保钾细菌肥的质量的研究。试验选用钾细菌P1为出发菌株 ,经紫外线、硫酸二乙酯交替处理 ,筛选出一株产菌量和芽孢量较高的突变株PE -R。用该菌株进行了发酵各因素的试验确定了最佳发酵条件 :接种量 2 0 %、淀粉浓度 1.2 %、氮源浓度 0 .3%、发酵初始pH7.0 - 7.2、发酵后期温度 37℃、发酵后期风量 1:1.2。试验结果表明 ,采用该工艺进行工业化生产 ,发酵菌量从 2 0× 10 10 个 /ml增加到 4 5× 10 10 个 /ml,芽孢生成量从 4 5 %增加到 95 % ,发酵时间从 2 4h缩短到 15h ,制成菌肥 1年后检测活菌保留率仍在 93 6 %。     

L-苹果酸产生菌F-871变株合成延胡索酸酶的研究*

温特曲霉AspergilluswentiiF-871是高效转化延胡索酸为L-苹果酸的变株,通过对其合成延胡索酸酶的因素进行研究,筛选了培养基主要营养元素有L-精氨酸、酪氨酸、丝氨酸、天冬氨酸、苏氨酸、赖氨酸及相应含量丰富的酵母粉、黄豆饼粉和玉米浆。该变株生长阶段条件为:pH6.0,温度2830℃,培养时间为24h,酶合成阶段最适条件:接种量15%,初始pH6.8,培养温度2830℃,装量8090ml/500ml,酶合成周期40h。在优化的培养条件下,F-871变株延胡索酸酶合成水平可达156.33u/g,100ml发酵液中的酶可将延胡索酸转化为L-苹果酸32.06g,比国内一步发酵法产酸88.5%提高约4倍,转化率由85%提高到106.87%,发酵周期由90h缩短至57h。     

L-苹果酸产生菌F-871变株合成延胡索酸酶的研究

温特曲霉AspergilluswentiiF-871是高效转化延胡索酸为L-苹果酸的变株,通过对其合成延胡索酸酶的因素进行研究,筛选了培养基主要营养元素有L-精氨酸、酪氨酸、丝氨酸、天冬氨酸、苏氨酸、赖氨酸及相应含量丰富的酵母粉、黄豆饼粉和玉米浆。该变株生长阶段条件为:pH6.0,温度2830℃,培养时间为24h,酶合成阶段最适条件:接种量15%,初始pH6.8,培养温度2830℃,装量8090ml/500ml,酶合成周期40h。在优化的培养条件下,F-871变株延胡索酸酶合成水平可达156.33u/g,100ml发酵液中的酶可将延胡索酸转化为L-苹果酸32.06g,比国内一步发酵法产酸88.5%提高约4倍,转化率由85%提高到106.87%,发酵周期由90h缩短至57h。     

酿酒酵母突变株J-X25胞内合成GSH的研究

以筛选得高产谷胱甘肽(GSH)产生酵母甲硫氨酸缺陷型变株J-X25为试验菌株。对其培养条件进行研究,结果表明：发酵培养基的最适初始pH值为6.0、最佳发酵温度为30℃、最佳装液量为100ml/500ml、接种量10%、摇床转速为220r/min。在酵母细胞培养到对数期,加入过氧化氢刺激细胞发生应激反应和乳酸钠作为表面活性剂改变细胞通透性,GSH总量达到0.253g/L,比不添加两者情况下的GSH产量高出52％。结果表明优化培养条件后,J-X25胞内积累GSH比出发株提高79％。     

黑曲霉产菊粉酶的发酵条件优化及诱变育种

对1株黑曲霉产菊粉酶的发酵条件进行了研究,确定了优化的发酵条件为菊粉2%,酵母膏2%,(NH4)H2PO40 5%,Na Cl0 5%,MgSO4·7H2O0 05%,ZnSO4·7H2O0 01%,初始pH6 5,接种量1 6%,装液量30ml,发酵5d,菊粉酶活最高可达26U/ml,I/S为1 15。经Co60诱变筛选出1株突变株C-32,在相同的发酵条件下菊粉酶活提高30%,I/S不变。     

L-苹果酸产生菌F-871变株合成延胡索酸酶的研究

温特曲霉Aspergillus wentii F-871是高效转化延胡索酸为L-苹果酸的变株,通过对其合成延胡索酸酶的因素进行研究,筛选了培养基主要营养元素有L-精氨酸、酪氨酸、丝氨酸、天冬氨酸、苏氨酸、赖氨酸及相应含量丰富的酵母粉、黄豆饼粉和玉米浆。该变株生长阶段条件为：pH6．0,温度2830℃,培养时间为24h,酶合成阶段最适条件：接种量15％,初始pH6．8,培养温度2830℃,装量8090ml／500ml,酶合成周期40h。在优化的培养条件下,F-871变株延胡索酸酶合成水平可达156．33u／g,100ml发酵液中的酶可将延胡索酸转化为L-苹果酸32．06g,比国内一步发酵法产酸88．5％提高约4倍,转化率由85％提高到106．87％,发酵周期由90h缩短至57h。     

巨大芽孢杆菌（Bacillus megaterium）突变株Bn,B5的生物学特性及在Vc二步发酵中的伴生作用

对选出的巨大芽孢杆菌突变株Bn,B5进行了生物学特性及发酵条件的研究,发现它们具耐低pH和抗高浓2KGA特性.可促进氧化葡萄糖酸杆菌生长,使其延迟期缩短,产酸增加.适宜的通气量下,摇瓶糖酸转化率提高10%～14%;当发酵pH为6.2～6.6时,转化率提高20%～30%.     

一株病原拮抗野生菌株的筛选、鉴定及其发酵工艺优化

为获得广谱抗菌功能野生菌株并提高其发酵产物中抗菌物质的含量。采用管碟法和菌丝生长速率法筛选功能菌株,ITS序列分析鉴定功能菌株,通过响应面法和正交设计优化发酵生产抗菌物质的工艺。筛选到一株强效、广谱抗菌功能菌株,鉴定为Cerrena sp.,其发酵产物对金黄色葡萄球菌、大肠杆菌、白色念珠菌、枯草芽孢杆菌和水稻纹枯病菌有显著拮抗作用。该菌株的摇甁发酵配方及培养条件为：马铃薯13.99 g/L,蔗糖 41.58 g/L,VB1 0.027 g/L,麸皮7 g/L,KH₂PO₄ 2 g/L,MgSO₄·7H₂O 2 g/L;摇床温度28 ℃、发酵周期10 d、种龄4 d、接种量8%、初始pH为5.0、装液量110 ml/250 ml。该菌株有明显抑菌活性,发酵工艺优化后抗菌活性提高了30.37%,为该菌株今后的应用、抗菌剂的分离提纯和产业化提供了实验依据。     

产核黄素阿舒假囊酵母的诱变效果及添加小米、肌醇的增产效果

用亚硝基胍和紫外线对产核黄素阿舒假囊酵母2.1197进行诱变处理,其核黄素发酵单位从372μg/ml提高到4660—6492μg/ml,提高幅度为25.3—74.5%;用紫外线和氮激光处理,提高62.3%;用红外线二氧化碳激光处理,提高39.7%。在该菌摇瓶发酵培养基中添加小米粉1%,发酵单位提高9.78%,添加1%小米粉和10μg/ml的肌醇,发酵单位提高16.84%。     

Semi-pilot scale studies on citric acid fermentation by a gamma-ray induced mutant of Aspergillus niger

Summary Semi-pilot scale production of citric acid was investigated with a gamma-ray induced mutant (HB3) of Aspergillus niger using 500, 1000 and 1500 ml medium in 51 fermentation jars. Yield of citric acid was found to be seven-fold higher compared to the parent in 1000 ml medium and the corresponding increase was two-fold in the 500 ml medium. With 1500 ml/fermentation jar the yield was low with both the parent and the mutant strain though the mutant gave higher yield compared to the parent.     

螺旋霉素产生菌的推理选育

螺旋霉素(spiramycin,以下简称SPM)为十六元大环内酯类抗生索〔r〕。内酯环C－3位上羟基酰化侧链片段构成SPM不同组分。C－3位上羟基酰化酶的合成受葡萄糖的诱导．该诱导作用受丁酸的抑制〔2〕。SPM的生物合成受高浓度铵离子的抑制。而支链氨基酸,如缬氨酸,经分解代谢可产生乙酸、丙酸、丁酸〔S〕。作为SPM生物合成的前体。本文根据SPM的生物合成途径,采用诱变和耐L－缬氨酸、耐α－氨基丁酸相结合的筛选方法．获得了高产菌株。     

1,3-丙二醇产生菌的诱变育种及培养基优化

以实验室自然筛选的克雷伯氏杆菌(Klebsiella sp.)为出发株,采用紫外诱变及亚硝基胍和超声波协同处理获得一株1,3-丙二醇高产突变株。在摇瓶发酵中,其产1,3-丙二醇产量由17.39 g/L提高到24.11 g/L,提高38.64%。变异株经10次传代培养,发酵能力稳定。对发酵培养基成分进行了优化,优化后1,3-丙二醇产量为30.05g/L,为优化前的1.25倍。     

脂肪酶产生菌的选育及产酶条件的优化

A lipase-producing bacterium strain was isolated from soil and was identified as Pseudomonas sp.. Its lipase yield was improved 2.25-fold by combined beatment of UV irradiation and NTG. The lipase fermentation condition for the mutant strain was optimized with Plackett-Burman design and Response Surface Analysis(RSA), and the formula of the optimum medium suitable for industrial scale fermentation was thereby established. A maximum yield of 87.5 U/ ml was obtained.     

Effect of cysteine on methionine production by a regulatory mutant of Corynebacterium lilium

The production of methionine by submerged fermentation using a mutant strain of Corynebacterium lilium was studied to determine suitable conditions for obtaining high productivity. The mutant strain resistant to the methionine analogues ethionine, norleucine, methionine sulfoxide and methionine methylsulfonium chloride produced 2.34 g l(-1) of methionine in minimal medium containing glucose as carbon source. The effect of cysteine on methionine production in a 15 l bioreactor was studied by supplementing cysteine intermittently during the course of fermentation. The addition of cysteine (0.75 g l(-1)h(-1)) every 2 h to the production medium increased the production of methionine to 3.39 g l(-1). A metabolic flux analysis showed that during cysteine supplementation the ATP consumption reduced by 20%. It also showed that the increase in flux from phosphoenol pyruvate to oxaloacetate leads to higher methionine production. Results indicate that controlling the respiratory quotient close to 0.75 will produce the highest amount of methionine and that regulatory mutants also resistant to analogues of cysteine would be better methionine over producers.     

产共轭亚油酸菌株的筛选及其等离子体诱变

本实验从东北酸菜汁中筛选获得了一株产共轭亚油酸能力较好的菌株SC-05,特别是气相色谱分析结果表明,合成的CLA产物构型为单一构型,仅为c9, t11-18：2-CLA,经鉴定该菌为乳杆菌属Lactobacillus sp.。菌株SC-05经低温等离子体处理后,筛选出突变株A-08,其共轭亚油酸产量达到119.24μg/mL,比出发菌株增加了83.0%,且突变株A-08具有较好的遗传稳定性。     

Strain improvement and up scaling of phytase production by <Emphasis Type="Italic">Aspergillus niger</Emphasis> NCIM 563 under submerged fermentation conditions

Combination of physical and chemical mutagenesis was used to isolate hyper secretory strains of Aspergillus niger NCIM 563 for phytase production. Phytase activity of mutant N-1 and N-79 was about 17 and 47% higher than the parent strain. In shake flask the productivity of phytase in parent, mutant N-1 and N-79 was 6,181, 7,619 and 9,523 IU/L per day, respectively. Up scaling of the fermentation from shake flask to 3 and 14 L New Brunswick fermenter was studied. After optimizing various fermentation parameters like aeration, agitation and carbon source in fermentation medium the fermentation time to achieve highest phytase activity was reduced considerably from 14 days in shake flask to 8 days in 14 L fermenter. Highest phytase activity of 80 IU/ml was obtained in 1% rice bran–3.5% glucose containing medium with aeration 0.2 vvm and agitation 550 rpm at room temperature on 8th day of fermentation. Addition of either bavistin (0.1%), penicillin (0.1%), formalin (0.2%) and sodium chloride (10%) in fermented broth were effective in retaining 100% phytase activity for 8 days at room temperature while these reagents along with methanol (50%) and ethanol (50%) confer 100% stability of phytase activity at 4°C till 20 days. Among various carriers used for application of phytase in feed, wheat bran and rice bran were superior to silica and calcium carbonate. Thermo stabilization studies indicate 100% protection of phytase activity in presence of 12% skim milk at 70°C, which will be useful for its spray drying.     

内切葡聚糖酶高产菌株的诱变选育

【目的】建立里氏木霉(Trichoderma reesei)高产突变菌株的快速筛选方法,选育出高产内切葡聚糖酶的突变株。【方法】对里氏木霉T306菌株的初筛培养基进行优化,建立快速筛选方法;通过紫外诱变手段选育内切葡聚糖酶高产突变菌株,并对突变菌株的产酶培养基进行优化。【结果】在初筛培养基中添加浓度为0.1%(W/V)的乳糖、蛋白胨及脱氧胆酸钠有利于菌株的筛选。诱变后筛选出菌落形态发生明显变化的内切葡聚糖酶高产突变株0516,其羧甲基纤维素酶活力(CMC酶)较出发菌株提高了38.9%。其产酶培养基经优化后,得到最适碳、氮源分别为:乳糖1.50%、硫酸铵0.14%、尿素0.05%、蛋白胨0.10%,优化后CMC酶活力达64.2 U/mL,较优化前提高了2.3倍。【结论】建立了里氏木霉高产突变菌株的快速筛选方法,通过紫外诱变育种获得了产内切葡聚糖酶能力高且遗传稳定的突变株0516。