高生物量富铁酵母菌的选育及其发酵条件的研究

对402株不同种属的酵母菌株进行初筛、复筛,筛选到一株生物量较高的二倍体菌株ZY-46(Saccharomyces cerevisiae)和一株铁富集量较高的二倍体菌株ZY-173(Saccharomyces kluyveri)。然后以它们为出发菌,分别进行单倍体分离、硫酸二乙酯(DES)诱变,并通过原生质体融合,得到一株高生物量富铁酵母融合菌株ZYF-15。在优化的发酵条件下,该融合菌株生物量可达11．2g／L,细胞铁含量达24．5mg／g干细胞,细胞总铁含量分别比原始亲株ZY-46和ZY-173提高了2．6倍和1．9倍。     

富锌酵母的选育及培养条件研究

对402株不同种属的酵母菌株进行了出筛、复筛、单倍体分离、诱变,从亲株Y-6-16-18（a met）accharomyces cerevisiae）和Y378-4-15（α-leu）（Sacharomyces kluyveri）的杂交菌株中选育到一株富锌酵母菌株（编号为ZGH374）。并初步优化了发酵条件：培养基为80g/L糖浓度的麦芽汁、10g/L蛋白胨、锌添加量400μg/mL,pH 6.0,装液量40mL/250mL三角瓶,接种量10%（V/V）,培养起始添加锌盐,培养时间30h。在优化的条件下,杂交菌株ZGH374的生物量（细胞干重）达到14.3g/L,细胞锌含量可达到9.3mg/g,锌总含量达到了133mg/L。     

高生物量富硒酵母的选育及发酵条件的研究

采用硒浓度耐性培养,从8株酿酒酵母中筛选驯化得到一株高生物量富硒酵母菌株NH-7,并对其发酵条件进了行优化。采取流加培养,在菌株稳定期,即18 h开始分批添加亚硒酸钠至总硒浓度为30μg/mL,控制乙醇浓度为0.4%和0.7%,分别得到20.14 g的高生物量(每升培养液中获得的酵母菌体干重)和82.52%的硒转化率。在60μg/mL总硒浓度的流加培养中,得到高达2 411μg/g的有机硒含量,每升发酵液菌体干重为18.83 g。     

红酵母类胡萝卜素形成的生理学研究

从数株红酵母中选出 1株产类胡萝卜素能力较强的红酵母RY 98(生物量、类胡萝卜素含量和产量分别为19.9g/L ,334 .8μg/ g和 6 .7mg/L) ;研究了该菌株产类胡萝卜素的最适营养与环境条件 ,获得了最佳的发酵生理学条件 :葡萄糖 40 g/L ,(NH4 ) 2 SO4 10 g/L ,酵母膏 3g/L ,蕃茄汁 2mL/L ,花生油 0 .5mL/L ,接种量 30mL/L ,初始pH 6 .0和通气量 (培养基装量 ) 4 0mL/ 2 5 0mL。在此初步优化的培养条件下 ,红酵母RY 98经 72h摇瓶发酵其生物量、类胡萝卜素含量和产量分别可达 2 6 .8g/L ,386 .9μg/ g和 10 .4mg/L ,依次比初筛中提高了 34 .7% ,15 .6 %和 5 5 .2 %。     

真姬菇液体发酵培养基筛选及硒富集条件优化

以真姬菇菌种为材料,采用液体摇瓶培养进行最适发酵培养基的优化,并探讨不同浓度亚硒酸钠和甘油的富硒效果。结果表明:真姬菇菌种最优液体发酵培养基为:马铃薯20%,麦麸滤液3%,可溶性淀粉3%,酵母粉0.15%,硫酸镁0.1%,磷酸二氢钾0.2%,最大生物量可达13.2 g/L;亚硒酸钠对菌丝生长有抑制作用,但可促进菌丝富硒,当其浓度为40μg/mL时,菌丝体的硒含量最大,达1.75μg/g,总体硒富集含量可达5.8μg/L发酵液;甘油对亚硒酸钠的总富硒含量有促进作用,当甘油浓度为50μg/mL时,菌丝总体硒富集含量最大,可达26.7μg/L发酵液。     

红酵母类胡萝卜素高产菌株的筛选及其发酵生理学条件研究

从数株红酵母中选出了1株产类胡萝卜素能力较强的红酵母RY－98（生物量,类胡萝卜素含量和产量分别为19．9g/L,334.8ug/g和6．7mg/L);研究了该菌株产类胡萝卜素的最适营养与环境条件,获得了最佳的发酵生理学条件;葡萄糖40g/L,(NH4)2SO4 10g/L,酵母膏3g/L,蕃茄汁2mL/L,花生油0.5mL/L,接种量30mLL初始pH6.0和通气量（培养基装置040ml/250mL:,在此初步优化的培养条件下,红酵母RY－98经72小时摇瓶发酵其生物量,类胡萝卜素含量和产量分别可达26．8g/L,386.9ug/g和10．4mg/L,依次比初筛中提高了34．7％,15．6％,和55．2％。     

高富硒酵母的筛选及富硒强化研究

富硒酵母在医学保健和制药产业中有着重要的应用,高富硒酵母是富硒酵母产品开发的基础。以新疆富硒土壤为样品来源,通过马丁培养基的初筛和富硒能力的复筛,筛选到一株富硒量相对较高的黏红酵母菌株Rhodotorula glutinis X-20。优化了菌株的富硒培养条件,结果显示,利用2%甘油为碳源可显著提高酵母中硒的累积,培养12 h时添加50μg/m L的亚硒酸钠能够保证酵母较高的硒累积量和生物量,添加0.4 mmol/L的磷酸盐可以有效地促进硒的转运和累积,最终使Rhodotorula glutinis X-20富硒量达到5 009μg/g。     

铁和镍对光合细菌生长和产氢的影响

基于金属元素在生物体功能发挥中的作用以及它们参与光合细菌光合放氢的重要性,着重进行了铁和镍对沼泽红假单胞菌（Rhodopseudomonas palustris）Z菌株和一株红杆菌（Rhodobactersp.）细胞生长、光合放氢和光合色素合成影响的研究。结果表明,高浓度Fe3+可显著提高两菌株光放氢能力和生物合成能力,最适浓度的Fe3+可使其产氢能力分别达对照组的1.32倍和2.8倍,产氢得率分别为360.6mL/g和385.9 mL/g,生物量分别为对照组的1.42倍和1.54倍。9μmol/L Ni2+的添加可使两菌株产氢能力分别达对照组的1.48倍和1.96倍,产氢得率分别为429.7mL/g和456.3 mL/g。而当Ni2+浓度为12μmol/L时,两菌株的产氢活性受到不同程度的抑制,产氢得率分别降低46.7%和19.4%。在铁浓度相同时,添加6μmol/L Ni2+能明显促进两菌株的生长。而当Ni2+浓度大于6μmol/L时,细胞生长受到抑制。Fe3+和Ni2+对Rhodobactersp.菌株类胡萝卜色素有显著影响。研究结果显示, 426nm色素峰随铁浓度的增加和镍的添加而消失,同时,产氢活性提高。     

酿酒酵母by1.1b产D-(-)-扁桃酸脱氢酶的发酵条件优化

对一株产D-(-)-扁桃酸对映选择性脱氢酶的酿酒酵母菌(Saccharomyces cerevisiae sp.strain by1.1b)发酵产酶条件进行了优化.研究各种碳源、氮源及无机盐对产酶的影响,应用正交试验优化发酵培养基组成,结果为:蛋白胨60 g/L,麦芽糖30 g/L,MgSO4 0.5 g/L,ZnSO4 0.01 g/L,KCl 1.0 g/L.优化后酶产量提高了7.9倍(由2.56 U/mL增至20.21 U/mL).摇瓶培养最佳条件为:装液量40%,发酵pH 6.5,接种量10%,发酵温度30℃.考察了细胞生长及产酶的时间进程,最佳培养时间为25 h.     

酿酒酵母by1.1b产D-(-)-扁桃酸脱氢酶的发酵条件优化

对一株产D-(-)-扁桃酸对映选择性脱氢酶的酿酒酵母菌(Saccharomyces cerevisiae sp. strain by1.1b)发酵产酶条件进行了优化。研究各种碳源、氮源及无机盐对产酶的影响, 应用正交试验优化发酵培养基组成, 结果为: 蛋白胨 60 g/L, 麦芽糖 30 g/L, MgSO4 0.5 g/L, ZnSO4 0.01 g/L, KCl 1.0 g/L。优化后酶产量提高了7.9倍(由2.56 U/mL增至20.21 U/mL)。摇瓶培养最佳条件为: 装液量40 %, 发酵pH 6.5, 接种量10 %, 发酵温度30 ℃。考察了细胞生长及产酶的时间进程, 最佳培养时间为25 h。     

Preparation of selenium yeasts I. Preparation of selenium-enriched Saccharomyces cerevisiae.

Selenium (Se) is an essential micronutrient for human and animal organisms. Organic selenium complexes and selenium-containing amino acids are considered the most bioavailable. Under appropriate conditions yeasts are capable of accumulating large amounts of trace elements, such as selenium, and incorporating them into organic compounds. It has been found that introduction of water-soluble selenium salt as a component of the culture medium for yeasts produced by conventional batch processing results in a substantial amount of selenium being absorbed by the yeast. Using a culture medium supplemented with 30 microg/mL sodium-selenite added during the exponential growth phase results in selenium-accumulation in the range of 1200-1400 microg/g dried baker's yeast (Saccharomyces cerevisiae) measured by ICP-AES method. In our previous studies it was shown that higher amounts of sodium-selenite in the culture medium have a strong inhibitory effect on the growth of this yeast. As a consequence of variations in cultivation conditions we obtained selenium yeast with different inorganic selenium content. The most important parameters influencing incorporated forms of selenium are pH value and dissolved oxygen level in the culture medium, and depending on these the selenium consumption rate of the yeast. A 0.40-0.50 mg/g h-1 specific selenium consumption rate was found to be appropriate to obtain selenium-enriched bakers' yeast of a high quality. Under suitable conditions the undesirable inorganic selenium content of the yeast could be suppressed to as low as 5-6% at the expense, however, of approximately a 20% decrease in the final biomass.     

Preparation of selenium yeasts I. Preparation of selenium-enriched Saccharomyces cerevisiae

Selenium (Se) is an essential micronutrient for human and animal organisms. Organic selenium complexes and selenium-containing amino acids are considered the most bioavailable.Under appropriate conditions yeasts are capable of accumulating large amounts of trace elements, such as selenium, and incorporating them into organic compounds. It has been found that introduction of water-soluble selenium salt as a component of the culture medium for yeasts produced by conventional batch processing results in a substantial amount of selenium being absorbed by the yeast.Using a culture medium supplemented with 30 μg/mL sodium-selenite added during the exponential growth phase results in selenium-accumulation in the range of 1200–1400 μg/g dried baker's yeast (Saccharomyces cerevisiae) measured by ICP-AES method. In our previous studies it was shown that higher amounts of sodium-selenite in the culture medium have a strong inhibitory effect on the growth of this yeast. As a consequence of variations in cultivation conditions we obtained selenium yeast with different inorganic selenium content. The most important parameters influencing incorporated forms of selenium are pH value and dissolved oxygen level in the culture medium, and depending on these the selenium consumption rate of the yeast. A 0.40–0.50 mg/g h-1 specific selenium consumption rate was found to be appropriate to obtain selenium-enriched bakers' yeast of a high quality. Under suitable conditions the undesirable inorganic selenium content of the yeast could be suppressed to as low as 5–6% at the expense, however, of approximately a 20% decrease in the final biomass.     

Selection of a high-biomass, chromium-rich yeast strain and optimization of cultivation conditions

Saccharomyces cerevisiae LZ-53 was selected from 240 primary yeast strains from different genera and species, whose chromium (Cr) resistance and biomass were higher than others were. The highest biomass and Cr content of the strain was obtained in 30 h at 28°C and 200 rpm, when 20 ml of the culture in 250-ml shake flasks was grown in wort containing 1200 μg/ml Cr. The initial pH was adjusted to 6.0. The optimal inoculum volume was 10% (v/v). The Cr content of the cells was determined by neutron activation analysis. Under the optimized cultivation conditions, the Cr content reached 3248 μg/g. Journal of Industrial Microbiology & Biotechnology (2001) 27, 195–198. Received 28 January 2001/ Accepted in revised form 11 June 2001     

The effect of pulse electric field on accumulation of selenium in cells of Saccharomyces cerevisiae

Cultures of Saccharomyces cerevisiae were subjected to the effect of PEF (pulse electric field) and a source of selenium. The culture period after which yeast cells were subjected to PEF treatment was optimized, as was the duration of the exposure. Optimization of the nutrient medium composition in S. cerevisiae cultures resulted in an over 1.8-fold increase in selenium accumulation with relation to cultures on the initial substrate. Optimization of the pH value and of culture duration resulted in selenium accumulation increase by approximately 78%. A significant correlation was found between the accumulation of selenium in yeast cells and its concentration in the culture substrate. The highest accumulation of selenium in the biomass of yeast, approx. 240 microg/g d.m., was obtained after 15-min exposure to PEF on a 20-h culture. An approx. 50% higher content of selenium in cells was recorded, as compared with the control culture without the application of PEF.     

Biotransformation of vegetable and fruit processing wastes into yeast biomass enriched with selenium

Water extracts of cabbage, watermelon, a mixture of residual biomass of green salads and tropical fruits were used for yeast cultivation. These extracts contained from 1420 to 8900 mg/l of dissolved organic matter, and from 600 to 1800 mg/l of nitrogen. pH of the extracts was in the range from 4.1 to 6.4. Biomass concentration of yeast, Saccharomyces cerevisiae CEE 12 grown at 30 degrees C for 96 h in the sterilized extracts without any nutrient supplements was from 6.4 to 8.2 g/l; content of protein was from 40% to 45% of dry biomass. The yield was comparable with the yield of yeast biomass grown in potato dextrose broth. The biomass can be considered as the protein source. Its feed value was enhanced by incorporation of selenium in biomass to the level of 150 microg/g of dry biomass. Therefore, it was recommended to transform the extracts from vegetable and fruit processing wastes into the yeast biomass enriched with selenium.     

富铬蚁巢伞（鸡菌）液体培养(英文)

A species of mushroom, Termitomyces albuminosus, was cultured in liquid medium for production of chromium-enriched mycelium. The influence of chromium (Ⅲ ) on mycelial growth of T. albuminosus was investigated. An optimum medium composed of 5.6g/L yeast extract, 51.6g/L hydrolyzed rice, 2g/L KH2PO4, and 20mg/L chromium(Ⅲ ) with initial pH of 4.5 was obtained by using method of central composite design (CCD). After incubation of 84h, the maximal biomass of chromium-enriched mycelia reached 24.23g DMW(dried mycelial weight)/L with 272μg/g DMW chromium content in 500mL flasks containing 100mL medium with an inoculum of 8% on a shaker of 100r/min under an optimized cultivation condition at 28℃.     

沼泽红假单胞菌富硒发酵条件的研究

通过对沼泽红假单胞菌富硒发酵条件的研究,确定最优的富硒培养方式。采用单一因素变量法,利用微波消解与紫外分光光度法测定沼泽红假单胞菌在不同发酵条件下的生物量变化与富硒效果,确定其最佳培养方式。结果表明,最佳富硒培养条件为:培养温度30℃,环境硒浓度100μg/mL,培养基初始pH为7,硒添加方式应为梯度分次添加,添加时间应为培养周期的第3天、第4天。利用得到的富硒菌种,在最优条件下培养7 d后,测得环境硒浓度下降为10.9μg/mL,硒的转化率为89.1%,菌体富硒量可达到73.54 mg/g(干菌种),其中有机硒含量为97.1%。利用沼泽红假单胞菌生产有机硒具有可行性,富硒菌体可以作为动物饲料添加剂,也可以为人类提供富含有机硒的食品。在以后的实验中,将进一步进行验证和工业化应用。     

虎掌菌菌丝富硒培养的研究

为生产合适的硒源提供一种思路,以菌丝体生物量、含硒量、还原糖、氨态氮和蛋白质为指标,采用四因素三水平正交设计法优化虎掌菌的富硒发酵条件,探讨不同浓度的硒对虎掌菌固体培养菌丝生长和液体培养产生的生物组分的影响。结果表明,高浓度的硒抑制虎掌菌菌丝体生长;正交试验选择不同的衡量指标,由极差分析得出各因素的影响程度大小,结合证实试验得到以还原糖为指标的最优组合为葡萄糖6%(质量分数)、酵母浸膏3%(质量分数)、KH_2PO_4 0.1%(质量分数)、Na_2SeO_3 0.6 mmol/L,其菌丝体生物量和氨态氮含量较高。与对照相比,加硒后虎掌菌发酵液中氨态氮、还原糖和总糖含量显著增加(P0.05);当硒浓度为0.5 mmol/L时,氨态氮、还原糖和总糖含量均达到最高值。菌丝体生物量和可溶性蛋白质分别在硒浓度为0.2 mmol/L和0.4 mmol/L时达到最大值。虎掌菌富硒培养后,发酵液的营养成分含量会发生变化。     

利用原生质体诱变育种选育富硒能力强的酵母菌株

利用原生质体诱变育种技术选育富硒能力强的酵母菌株,从13株啤酒酵母中筛选出一株富硒量高的诱变出发菌株,采用溶壁酶进行破壁,确定了原生质体制备的最适条件为酶浓度1g／100mL,酶解处理时间为120min,原生质体形成率为95．2％,再生率为21．8％,诱变后筛选出富硒量为821mg／kg,酵母干菌体收获量为0．88g／100mL的酵母菌Al。