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.     

12C6+离子对阿维链霉菌累进辐照诱变效应研究

选用12C6+ 离子束对阿维链霉菌诱变选育高产菌株与原始菌株进行辐照诱变, 研究其累进辐照效应。实验结果表明,在辐照剂量为10 Gy时, 原始菌株比诱变高产菌株存活率高, 抗辐射能力强;辐照剂量高于30 Gy时,诱变高产菌株比原始菌株存活率高, 抗辐射能力强。原始菌株正突变率最高的辐照剂量为50 Gy, 致死率99.43%,正突变率最高, 达34.2%;对诱变高产菌株辐照剂量为30 Gy,致死率94.97%,正突变率最高, 达23.5% 。累进辐照效应降低了最佳辐照剂量。 Mutagenic effect on the mutant high producing strain ZJAV Y1 203 and the original strain ZJAV A1 irradiated by ion beam of 12C6+ have been investigated. The experimental results indicated that the original strain has higher survival rate and stronger resistance to radiation than mutant high producing Strain at dose of 10 Gy. The mutant high producing strain has higher survival rate and stronger resistance to radiation than the original strain at the dose higher than 30 Gy. The lethality was 97% and the highest rate of orthomutation was 34.2%, when ZJAV A1 was irradiated by 50 Gy 12C6+ beam. The lethality was 94.97% and the highest rate of orthomutation was 23.5% when ZJAV Y1 203 was irradiated by 30 Gy 12C6+ beam. The best radiation dose is decreased by progressivity irradiation.     

~(12)C~(6+)离子辐照对苜蓿M1代低温胁迫存活率、过氧化氢酶及过氧化物酶活性的影响

研究了不同剂量12C6+离子辐照对中兰1号、BC-04-477、塔城3种苜蓿M1代个体在低温胁迫下存活率、过氧化氢酶(CAT)及过氧化物酶(POD)活性的影响。在辐照剂量为400 Gy时,中兰1号低温胁迫组存活率、CAT活性比未辐照的对照分别提高了33.3%,56.3%,POD活性与未辐照的对照无差异;在辐照剂量为400 Gy时,BC-04-477低温胁迫组存活率、CAT及POD活性比未辐照的对照组分别提高了33.3%,69.2%,5.1%;塔城在辐照剂量为800 Gy时,低温胁迫组的存活率、CAT及POD活性比未辐照的对照组分别提高了25%,26%,22.8%。以上结果表明,12C6+离子辐照可以提高中兰1号、BC-04-477、塔城的低温环境的存活能力,提高苜蓿抗寒性能。     

阿佛曼链霉菌的~(12)C离子辐照效应及高产菌株选育

选用12C6+离子辐照诱变阿维菌素B1a产生菌ZJAV-A1,研究其诱变效应。实验结果表明,12C6+离子辐照剂量50Gy时致死率97%,正突变率最高可达到34.2%。通过12C6+离子诱变处理,结合平板培养基及斜面培养基的正突变菌株筛选,最终获得一株稳定性良好,阿维菌素B1a组分产量稳定在4460—4588μg/ml之间,较出发菌株提高11.1%—14.7%的突变株ZJAV-Y1-203。     

重离子束对北里链霉菌的诱变效应

考察不同剂量重离子束对北里链霉菌的致死及诱变效应, 可确定最有利于筛选高产菌株的重离子束辐照剂量。 利用不同剂量的重离子束辐照北里链霉菌孢子, 统计了存活率、 致死率、 正负突变率。 结果发现, 在5 Gy重离子辐照时北里链霉菌出现较高致死率, 其后随剂量升高, 致死率变化较平缓。 各辐照剂量下正负突变率相比较, 40 Gy时正突变率最高, 负突变率相对较低, 存活率为0.92%。 因此确定40 Gy是对北里链霉菌高产菌株筛选最有利的辐照剂量。 To define the optimum dose of heavy ion beams for selecting high productive strains, we should study mortality and mutation effects of Streptomyces kitasatoensis irradiated by heavy ion beams in diffe rent doses. In this research, spores of Streptomyces kitasatoensis were irradiated by heavy ion beams with different doses. And survival rate, mortality rate, positive mutation and negative mutation were analyzed statistically. The results showed that high mortality rate appeared from 5 Gy and then the mortality rate curve became gently. Compared the positive and negative mutations in different doses, highest positive mutation was obtained in 40 Gy, while the negative mutation was lower in this dose, and the survival rate was 0.92%. So we defined that optimum dose of heavy ions radiation for Streptomyces kitasatoensis selection was 40 Gy in this experiment.     

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%。     

Influence of 12C6+ ion irradiation on mutant avermitilis

The effects of ¹²C⁺⁶ ion irradiation on colony morphology and mycelia morphology, as well as on mutation rate have been studied in the B1a high-product strains (ZJAV-Y1-203) mutated by heavy ion irradiation and compared with that in the original strain (ZJAV-A-1). After irradiating the rate of a straw hat colony type having a high ability of producing B1a in ZJAV-Y1-203 strains was higher than that found in ZJAV-A-1 strains. When strains were cultured in a liquid medium for 24 hours, the mycelium becoming thinner could be observed in all of the irradiated ZJAV- Y1-203 groups, but only in the ZJAV-A-1 groups irradiated at the dose of 50 Gy or more. The early growth of mycelium was inhibited in the ZJAV- Y1-203 group irradiated with a high dose. The highest positive mutation rate (23.5%) of ZJAV - Y1 - 203 was reached at the lower dose of 30 Gy while the highest positive mutation rate of 34.2% in ZJAV-A-1 appeared at 50 Gy.