激光诱变选育果胶酶高产菌株

采用Ar＋、He－Ne、LD、YAP等激光辐照果胶酶产生菌，不同波长激光辐照均获高酶活菌株，其中从LD激光与He－Ne激光辐照组中选育出酶活达6×104单位／g的高产菌株（比对照组提高122％），并稳定地应用于生产．文中从黑曲霉抱子与DNA的吸收光谱以及电镜观察黑曲霉抱子的结果出发，对激光诱变机制进行了初步探讨．     

氦-氖激光诱变细菌细胞及原生质体选育果胶酶高产菌株

利用氦-氖激光(波长632.8nm,功率15mW)诱变果胶酶产生菌的菌体细胞,获得了突变株ZH-g,其酶活达到301 u·mL－1,是出发菌株的1.3倍.进一步用氦-氖激光照射ZH-g的原生质体,从而得到了高产突变株ZH-gA,其酶活达到357 u·mL－1,比ZH-g提高了18%,是出发菌株的1.6倍.该菌株在好氧和静置条件下均能良好产酶,经传代实验证明其产酶性能稳定.     

激光诱变选育果胶酶高产菌株

采用Ar⁺、He-Ne、LD、YAP等激光辐照果胶酶产生菌,不同波长激光辐照均获高酶活菌株,其中从LD激光与He-Ne激光辐照组中选育出酶活达6×10⁴单位/g的高产菌株(比对照组提高122%),并稳定地应用于生产.文中从黑曲霉抱子与DNA的吸收光谱以及电镜观察黑曲霉抱子的结果出发,对激光诱变机制进行了初步探讨.     

紫外激光等选育生淀粉糖化酶高产菌株及其应用

以紫外线、LiCl、5-氟尿嘧啶、HNO2为诱变剂,黑曲霉Sx为出发菌株,选育出生淀粉糖化酶酶活力提高12%的突变株黑曲霉S′x.制取黑曲霉S′x的原生质体,用He-Ne激光照射,经反复筛选得到突变株黑曲霉Sy,其酶活力较出发菌株提高51%,而酸性蛋白酶活力降低45%.并对其在农作物秸杆的应用进行了研究.     

紫外激光等选育生淀粉糖化酶高产菌株及其应用

以紫外线、LiCl、5氟尿嘧啶、HNO₂为诱变剂,黑曲霉Sx为出发菌株,选育出生淀粉糖化酶酶活力提高12%的突变株黑曲霉S’x制取黑曲霉S’x的原生质体,用HeNe激光照射,经反复筛选得到突变株黑曲霉Sy,其酶活力较出发菌株提高51%,而酸性蛋白酶活力降低45%并对其在农作物秸杆的应用进行了研究.     

辐照前处理木质纤维素的研究进展

本文对几种主要的辐照前处理木质纤维素方法的效果和特点进行较为全面的比较和总结,旨在阐述微波、紫外线、γ射线、X射线、电子束、离子束等方式辐照前处理木质纤维素对其结构改变的机理尤其重点探讨了重离子辐照前处理引起木质纤维素同质异形体（I_α → I_β）的转换机制。木质纤维素结晶度与酶消化率呈强相关性,重离子辐照前处理可使木质纤维素中木质素和半纤维素部分破坏,导致其相对结晶度增加,从而增强了纤维素酶与木质纤维素的可及度,提高了酶解产率。因而,通过适当剂量的重离子辐照前处理可以显著提高酶对木质纤维素的生物转化效率和还原糖的产量,这为辐照前处理提高木质纤维素的综合利用提供了理论指导。     

紫外、氦氖激光等复合诱变产果胶酶细菌ZH1的研究

ZH₁菌株是从自然界分离筛选而来的,该菌株产果胶酶的最适pH为7.0;最适温度33℃;培养36h达产酶高峰,酶活力为73.5μ/mL,将ZH₁作为出发菌株,经紫外线诱变、硫酸二乙酯诱变,亚硝基胍和紫外线复合诱变及氦氖激光等多次反复诱变,选育得到一株产果胶酶性稳定且酶活明显提高的突变株ZHg,其酶活为301m/mL,比出发菌株ZH₁产果胶酶能力提高3.1倍.     

纤维素酶水解老化纸张中纤维素物质的酶活力分析

应用3,5-二硝基水杨酸测定法,分析了纤维素酶水解老化纸张中纤维素物质的酶活力影响因素,根据酶活力确定了酶解反应条件。pH=4.8,温度为50℃,酶解时间为60min,加酶量为10kU/mL条件下,经纤维素酶水解后测得的还原糖量呈现一定的趋势,可以此来初步判定纸张纤维的老化程度。     

C离子束诱变产生甜高粱汁酒精酵母高产菌株的研究

利用100 MeV/u C离子束对高产酒精酵母菌株进行了辐照诱变的研究。 采用红四氮唑作为筛选指示剂, 得到了5株产酒能力有所提高的突变酵母菌。 利用甜高粱汁的发酵结果表明, T4突变菌株的产酒精能力比原始出发菌株提高了18.6%, 且发酵液中的残糖含量也有所降低。 随后对T4菌株在甜高粱汁中的最适宜发酵条件做了初步探索, 结果表明: 最适发酵温度和pH值分别为30 ℃和4.5。 通过10 l发酵罐的验证试验表明: 在同样发酵条件下, T4菌株的发酵率和产酒精能力都比原始出发菌株提高了12%。 Five mutants with high ability of producing alcohol were selected out by using TTC as an indicator after irradiation of the alcohol yeast with 100 MeV/u carbon ions. The fermentation experiment in sweet sorghum juice showed that the alcohol production ability of mutant T4 strain increased 18.6% compared to the control strain. The residual sugar content in the juice was decreased too. After that, the optimum fermentation conditions of the T4 strain in sweet sorghum juice were investigated. The results showed that the optimum temperature and pH value for fermentation were 30 ℃ and 4.5, respectively. The verification experiment was fermented in a 10 l bio reactor and the obtained data indicated that the fermentative rate and the ability of producing alcohol in T4 strain was higher than that in the control strain under the same fermentation condition.     

12C6+离子辐照对阿维链霉菌代谢效应研究

对12C6+ 离子辐照诱变高产阿维链霉菌株ZJAV-Y1-203 和原始菌株ZJAV-A-1 的摇瓶发酵pH 值、菌体浓度、碳源和氮源代谢进行了测定,研究了12C6+ 辐照对阿维链霉菌的代谢效应。在发酵前期(48h),原始菌株发酵液pH 值低于突变菌株;在发酵96~196 h,诱变高产菌株ZJAV-Y1-203 繁殖快,生长旺盛,N的利用率高;菌体浓度大于原始菌株的浓度,且发酵液的pH 稳定,菌体处于代谢相对更稳定期;在发酵144~240 h,诱变高产菌株ZJAV-Y1-203 对糖源消耗低于原始菌株ZJAV-A-1。这些结果表明,12C6+ 离子辐照对阿维链霉菌代谢影响有利于阿维菌素合成。pH value, mycelium concentration, carbon source and nitrogen metabolism in flask fermentation of the mutant high-producing strain ZJAV-Y1-203 and the original strain ZJAV-A1 have been investigated, in order to show the metabolic effect of avermitilis irradiated by ion beam of 12C6. In early stage (48 h) of the fermentation, pH value of the original fermentation was lower than that of the mutant strains. In 96~196 h of fermentation, the nitrogen utilization in the strains ZJAV-Y1-203 was higher than that in the original strains, its reproductive was fast, and its growing was vigorous. The mycelium concentration of ZJAV-Y1-203 was greater than the original strain, and the pH value of fermentation were stable, so its metabolism was relatively more stable. In 144240 h of fermentation, the strain ZJAV-Y1-203 on sugar consumption was less than the original strains. The effect of 12C6 ion irradiation on metabolism of Streptomyces avermitilis is conducive to the synthesis of avermectin.     

重离子束12C6+累进辐照诱变柠檬酸菌株选育研究

应用大剂量重离子束12C6+ 对菌株H3001 进行二次辐照选育,对初选获得的高产柠檬酸菌株进行摇瓶发酵试验及10~ 100 L 中试发酵罐试验,采用酸碱中和法测定发酵液中柠檬酸的含量。结果表明:当二次重离子12C6+剂量为857.8 Gy 时,致死率和正突变率达到最大值,分别为94.5% 和8%。通过摇瓶发酵试验,最终获得一株高产柠檬酸菌株hw317,控制该菌株发酵周期为60 h,柠檬酸酸度能达到19.2±0.2%。Heavy 12C6+ ion beams in various high doses were employed to irradiate H3001 strain for screening Aspergillus niger strain for hyper citric acid production. Three high-yield strains were obtained after shaker fermentation test. Among the three strains, the strain hw317 was implemented shaker fermentation for stability test and 10~100 L pilot fermentation tank for citric acid productive maximization. Acid-base neutralization method was applied to determinate the content of citric acid in fermented liquid. The results showed that: when the secondary heavy ion 12C6+ dose was 857.8 Gy, both of the fatality rate (94.5%) and the positive mutation rate (8%) were highest. Through the shaker fermentation tests and 10 ~ 100 L pilot fermentation test, one strain hw317 was screened and obtained for hyper citric acid production. Consequently, the final citric acid acidity can reach up to 19.2±0.2% with controlling fermentation cycle for 60 h.     

高压对米曲霉理化性质影响及诱变的研究

 在100~400 MPa压力、保压20 min的条件下处理酱油酿造菌种——米曲霉,研究高压对米曲霉存活率、形态特征、生理性质、蛋白酶、淀粉酶活性等的影响,并诱变筛选优良菌株。结果表明：高压对米曲霉的存活率、形态特征有明显的影响;压力对蛋白酶及淀粉酶活性的影响也有特异的规律,即在一定压力范围内（0.1~200 MPa）蛋白酶的活性随着压力的增加而减小,但随着压力的进一步升高（200~400 MPa）蛋白酶的活性又逐渐增强,在300 MPa时超过对照组,400 MPa时蛋白酶的活性达到最高值;淀粉酶在0.1~100 MPa时活性下降,在200 MPa时其平均的糊化和糖化活性最强、活力最高,当压力升高活性又开始降低,400 MPa时几乎又回到对照值。另外,高压处理后获得一株理想的变异菌株HP300a：生长速度快、孢子数量多、蛋白酶活力高,且不易被杂菌污染。其成曲的几项主要指标均优于对照株,酿出酱油的几项主要指标也明显优于对照株。为利用高压诱变筛选米曲霉优良菌种、提高酿造酱油产品的产量及质量提供了理论依据,并发现了高压处理米曲霉引起其蛋白酶及淀粉酶活性改变的特殊规律。     

He-Ne激光在异种间原生质体融合中的应用

用632.8nm 12mW的氦氖激光及聚乙二醇（PEG）复合诱导融合枯草芽孢杆菌与黑曲霉原生质体.为了提高枯草芽孢杆菌的糖化酶产酶能力,对两亲本在融合中采用不同的激光照射时间,选育出的融合子与枯草芽孢杆菌亲本相比,糖化酶酶活提高了2倍,并通过酯酶同工酶谱分析对融合子进行了鉴定,结果表明:融合子遗传性状与亲本相比发生了显著变化,通过传代培养,融合子具有良好的遗传稳定性.     

Brewing characteristics of piezosensitive sake yeasts

Application of high hydrostatic pressure (HHP) treatment to food processing is expected as a non-thermal fermentation regulation technology that supresses over fermentation. However, the yeast Saccharomyces cerevisiae used for Japanese rice wine (sake) brewing shows high tolerance to HHP. Therefore, we aimed to generate pressure-sensitive (piezosensitive) sake yeast strains by mating sake with piezosensitive yeast strains to establish an HHP fermentation regulation technology and extend the shelf life of fermented foods. The results of phenotypic analyses showed that the generated yeast strains were piezosensitive and exhibited similar fermentation ability compared with the original sake yeast strain. In addition, primary properties of sake brewed using these strains, such as ethanol concentration, sake meter value and sake flavor compounds, were almost equivalent to those obtained using the sake yeast strain. These results suggest that the piezosensitive strains exhibit brewing characteristics essentially equivalent to those of the sake yeast strain.     

蛋白饲料酵母高产菌株的选育及发酵工艺优化研究

饲料中的蛋白含量,是衡量饲料优劣的主要指标,而决定饲料蛋白含量高低的主要因素包括:菌体蛋白含量(酵母菌、乳酸菌等)及饲草中自身蛋白含量;其中饲料酵母菌株蛋白含量直接决定了饲料蛋白的含量,因此获得优良的饲料酵母菌株成为关键。本研究利用辐射能量为80 MeV/u的12C6+重离子束对出发饲料酵母菌种NJ3236 (蛋白质量分数为40.64%)进行辐照诱变,并对辐照后的菌种进行初筛、复筛得到高蛋白含量菌株100G-2,该菌株较NJ3236蛋白含量提高了10.08%。利用响应面分析法对发酵培养基进行优化,通过优化得到的培养基的最优配比为:甜高粱汁20.95 g/L、玉米浆干粉18.17 g/L、硫酸镁1.60 g/L,在此条件下可溶性蛋白浓度达到1.381 mg/mL,较未优化前可溶性蛋白提高了8.7%。     

重离子诱变酵母培养基的优化与研究

利用12C6+离子诱变技术对面包酵母菌种进行诱变,得到一株粗蛋白含量达到55% 以上的菌株,借助Minitab16.0,采用Plackett-Burman 实验设计法及响应面分析法,对诱变后面包酵母菌发酵培养基的成分进行了优化,得到3 个最为显著的主要影响因子:葡萄糖、酵母抽提物和硫酸镁。利用最陡爬坡实验逼近最大响应区域后,利用Box-Behnken 实验设计及响应面分析法进行回归分析。通过求解回归方程,得到优化发酵的条件为,葡萄糖:11.03 g/L、酵母抽提物:6.53 g/L、硫酸镁:5.59 g/L。面包酵母生物量为4.84 g/L,相比未进行优化时的生物量提高了15%。A mutant bread yeast strain with high protein content of 55% was gained by use of 12C6+ ions. The MINITAB 16.0 software, Plackett-Burman experimental design and response surface methodology were applied to optimize the culture medium for the irradiated yeast. The most important three factors which influenced the culture results were identified as glucose, magnesium sulphate and yeast extract. The path of the steepest ascent was undertaken to approach the optimal region of the three significant factors. Box-Behnken design and response surface methodology were used for the regression analysis. Finally, the optimal fermentation conditions were identified as glucose 11.03 g/L, yeast extract 6.53 g/L and magnesium sulphate 5.59 g/L by the regression analysis. It was found that the biomass of the bread yeasts reached 4.84 g/L and increased by 15% compared to original conditions.     

Sonication of chitin-glucan, preparation of water-soluble fractions and characterization by HPLC

A water-insoluble chitin-glucan complex, isolated from the mycelium of Aspergillus niger, was swollen in various aqueous media and treated subsequently by high-energy sonication. The concentration of the resulting water-soluble polysaccharide fractions was dependent on the swelling medium, the amount of the chitin-glucan complex in the suspension, and on the time of sonication. The yields of water-soluble chitin-glucan were within the range 13.6 to 24.4% relative to the mass of the original chitin-glucan. The nitrogen content obtained for the samples of water-soluble chitin-glucan indicated a higher content of chitin (3.45% of nitrogen in high-molecular fraction) than in the original water-insoluble chitin-glucan sample (1.8%). The distribution of the molecular weights of the water-soluble fractions prepared was determined by high-performance liquid chromatography.