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。     

休哈塔假丝酵母乙醇发酵性能的研究

木糖的高效发酵是制约纤维素燃料乙醇生产的技术瓶颈之一,高性能发酵菌种的开发是本领域研究的重点。以木糖发酵的典型菌株休哈塔假丝酵母为材料,研究氮源配比、葡萄糖和木糖初始浓度、葡萄糖添加及典型抑制物等因素对其木糖利用和乙醇发酵性能的影响规律。结果表明,硫酸铵更适宜于木糖和葡萄糖发酵产乙醇。在摇瓶振荡发酵条件下,该酵母可发酵164.0 g/L葡萄糖生成61.9 g/L乙醇,糖利用率和乙醇得率分别为99.8%和74.0%;受酵母细胞膜上转运体系的限制,对木糖的最高发酵浓度为120.0 g/L,可生成45.7 g/L乙醇,糖利用率和乙醇得率分别达到94.8%和87.0%。休哈塔假丝酵母发酵木糖的主要产物为乙醇,仅生成微量的木糖醇;添加葡萄糖可促进木糖的利用;休哈塔假丝酵母在葡萄糖发酵时的乙酸和甲酸的耐受浓度分别为8.32和2.55 g/L,木糖发酵时的乙酸和甲酸的耐受浓度分别为6.28和1.15 g/L。     

S-腺苷甲硫氨酸和谷胱甘肽联产发酵的环境条件

在5 L发酵罐中,研究pH、搅拌转速和温度等环境条件对产朊假丝酵母CCTCC M209298联产发酵合成S-腺苷甲硫氨酸(SAM)和谷胱甘肽(GSH)的影响,发现酵母细胞生长、SAM和GSH合成各自需要最适的pH、搅拌转速和培养温度。以SAM和GSH联产量最大化为目标,获得了较为合适的联产发酵条件:pH 5.0,搅拌转速350 r/min,温度30℃。在此环境条件下,结合不低于35%的溶氧体积分数,分批培养产朊假丝酵母24 h,最终SAM和GSH联产产量可达到579.6 mg/L。     

秦巴山区富硒蛹虫草有效成分及硒存在形态分析

为探究秦巴山区富硒蛹虫草有效成分及硒存在形态,以秦巴山区蛹虫草CM-1518为研究对象,研究不同质量分数亚硒酸钠(0～500 mg/kg)对蛹虫草生长发育的影响,并对其有效成分及硒存在形态进行分析。试验结果表明当亚硒酸钠质量分数为100 mg/kg时,蛹虫草鲜质量最高,为293.41 g/盒。当亚硒酸钠质量分数为200 mg/kg时,虫草素、虫草酸含量最高,分别为1.06 mg/g和2.10 mg/g,表明硒与虫草素和虫草酸可协同增效,但虫草多糖含量变化规律不明显,亚硒酸钠的添加不利于腺苷的合成积累。经计算,富硒蛹虫草中有机硒所占百分比均高达99.9%,低浓度的亚硒酸钠可促进可溶性蛋白和可溶性多糖中硒的合成,但高浓度的硒却降低其合成,且富硒蛹虫草中可溶性多糖中硒含量高于可溶性蛋白硒含量。试验表明适宜浓度的亚硒酸钠可促进蛹虫草生长发育及有效成分合成积累。     

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。     

溶氧及pH对产朊假丝酵母分批发酵生产谷胱甘肽的影响

在7 L发酵罐中研究了溶氧和pH对产朊假丝酵母分批发酵生产谷胱甘肽的影响。结果表明,当葡萄糖浓度为30 g/L且通气量控制在5 L/min时,搅拌转速达到300 r/min即可满足细胞生长和谷胱甘肽合成对溶解氧的需求。不同pH控制方式对谷胱甘肽分批发酵的影响有较大差异。不控制pH时,细胞干重和谷胱甘肽产量比控制pH为55的发酵分别低27%和95%,且有50%的谷胱甘肽向胞外渗漏。研究了将pH控制在4.0、4.5、5.0、5.5、6.0和6.5的谷胱甘肽分批发酵过程,发现在pH 5.5时谷胱甘肽总产量最高。用前期研究建立的动力学模型模拟了不同pH (4.0～6.5)下的分批发酵过程,并从动力学角度解释了pH对细胞生长和谷胱甘肽合成的影响。     

产朊假丝酵母发酵生产谷胱甘肽的补料研究

为了实现产朊假丝酵母( Candida utilis WSH02-08)的高密度培养和提高谷胱甘肽 ( glutathione, GSH)产量,在分批发酵研究的基础上,考察了指数速率流加和DO-stat反馈流加对产朊假丝酵母合成GSH的影响.结果表明,采用指数速率流加可获得高细胞密度,但不利于GSH的合成,而采用DO-stat反馈流加较适宜细胞积累GSH.因此,提出并运用了指数速率-DO-stat组合流加策略,即采用指数速率流加实现细胞高密度培养,采用DO-stat反馈流加实现GSH的高积累,细胞量、GSH产量和胞内GSH含量分别为82.5 g/L、1120.6 mg/L和1.46%,实现了高细胞密度和高胞内GSH含量的相对统一.     

过氧化氢胁迫促进产朊假丝酵母合成谷胱甘肽

谷胱甘肽(GSH)在生物细胞抵御外界环境条件的刺激和胁迫时起到非常重要的作用。考察了不同时间不同浓度过氧化氢胁迫和过氧化氢连续胁迫对产朊假丝酵母合成GSH的影响, 发现低浓度过氧化氢的连续胁迫对GSH的合成有明显促进作用。进一步在发酵罐上应用了低浓度过氧化氢(36 mmol/L)持续胁迫策略, 最终GSH产量为922 mg/L, 胞内GSH含量为1.64%, 比对照分别提高了7%和35%。     

响应面法优化产朊假丝酵母培养及干燥工艺

本研究旨在优化产朊假丝酵母液体培养参数及干燥保护剂配方,提高产朊假丝酵母液体培养数量,制备出高复苏率的活性干酵母。以装液量、转速、温度和培养时间为影响因素,设计单因素试验,并采用Box-Behnken试验设计及响应面分析法优化产朊假丝酵母液体培养方案。在此条件下培养酵母,以L-谷氨酸钠、乳糖、脱脂奶粉为影响因素进行单因素保护试验,并采用响应面分析法优化干燥保护剂的组合。结果表明,产朊假丝酵母最佳培养条件为装液量45 mL/250 m L,转速200 r/min,温度30℃,培养时间24 h。在此条件下,培养的产朊假丝酵母数量可以达到9.34×10~8CFU/mL;最佳保护剂组合1%L-谷氨酸钠、12%脱脂奶粉、6%乳糖,经干燥后,产朊假丝酵母复苏率达到81.9%。此时活菌数7.65×10~8CFU/g,比未加保护剂组提高了3.83倍。     

过氧化氢胁迫促进产朊假丝酵母合成谷胱甘肽

谷胱甘肽(GSH)在生物细胞抵御外界环境条件的刺激和胁迫时起到非常重要的作用。考察了不同时间不同浓度过氧化氢胁迫和过氧化氢连续胁迫对产朊假丝酵母合成GSH的影响, 发现低浓度过氧化氢的连续胁迫对GSH的合成有明显促进作用。进一步在发酵罐上应用了低浓度过氧化氢(36 mmol/L)持续胁迫策略, 最终GSH产量为922 mg/L, 胞内GSH含量为1.64%, 比对照分别提高了7%和35%。     

Efficient Preparation of Selenium/Glutathione-Enriched Candida utilis and Its Biological Effects on Rats

The main purpose of this study was to prepare selenium/glutathione-enriched Candida utilis and investigate its effect on growth performance, antioxidant capacity, and immune response in rats. The preparation of the selenium/glutathione-enriched yeast was conducted using fed-batch culture for high cell density. The optimal culture conditions for increased intracellular organic selenium and glutathione contents were as follows: the concentrated medium was fed beginning at 12?h using a polynomial feeding strategy until a total glucose concentration of 150?g/l was reached, and sodium selenite was continuously added together with glucose to a total concentration of 60?mg/l. As a result, 81?% of sodium selenite was assimilated and transformed into organic selenium by C. utilis under optimal conditions, which in turn resulted in greater glutathione accumulation and lower malondialdehyde cellular content in the yeast. To investigate and compare the effects of the prepared selenized C. utilis and other dietary supplements, 40 female rats were divided into five groups of eight rats each, following a randomized block design. Experimental feeding was conducted for a period of 6?weeks. Selenium supplementation with inorganic selenium (sodium selenite) and organic selenium (selenized C. utilis) showed better results than the control and other groups supplemented with yeast with or without glutathione. The body mass of rats, selenium deposition, and oxidative enzymes activities in both serum and liver samples, and immunity responses were all significantly improved by selenium supplementation, and between the two sources, organic selenium was more effective than inorganic selenium.     

Selenium-enriched Candida utilis: Efficient preparation with l-methionine and antioxidant capacity in rats

Selenium-enriched Candida utilis has attracted much attention due to its expanding application in food and feed additives. The objective of this study was to efficiently prepare selenium-enriched C. utilis and to investigate the effects of the prepared yeast on antioxidant capacity in rats. A batch culture of selenium-enriched C. utilis was first carried out, and the addition of sodium selenite (Na₂SeO₃) after all glucose had been consumed was found to favor higher intracellular glutathione and organic selenium content. Moreover, l-methionine boosted yeast cell growth and glutathione biosynthesis, and prevented glutathione from leaking to the extracellular space that can be caused by Na₂SeO₃. We therefore developed a two-stage culture strategy involving supplementation with l-methionine and Na₂SeO₃ at separate culture phases to improve the performance of selenized C. utilis. Using this two-stage culture strategy, intracellular glutathione content reached 18.6 mg/g and 15.5 mg/g, respectively, in batch and fed-batch systems, and organic selenium content reached 905.2 μg/g and 984.7 μg/g, respectively. The effects of selenium-enriched C. utilis on the activities of antioxidant related enzymes in rats were investigated, and the prepared selenium-enriched C. utilis was shown to be an optimal dietary supplement for enhancing antioxidant capacity in rats.     

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。     

Performance,carcass characteristics and meat quality of Nellore cattle supplemented with supranutritional doses of sodium selenite or selenium-enriched yeast

The enrichment of meat with selenium is important to improve the intake of selenium by humans. The effects of supranutritional doses of sodium selenite or selenium-enriched yeast on performance, carcass characteristics and meat quality were evaluated using 63 Nellore cattle in a completely randomized design with two sources (sodium selenite and selenium-enriched yeast), three levels (0.3, 0.9 and 2.7 mg Se/kg DM) and control treatment (without addition of selenium). Final body weight (BW), average daily gain, dry matter intake and gain to feed ratio (G : F) at the end of 84 days of supplementation were not influenced by treatments (P>0.05). Values of pH, ribeye area, back fat thickness and marbling score were also not influenced by treatments (P>0.05). Dressing percentage was greater (P=0.02) in Nellore cattle supplemented with organic Se (58.70%) compared to animals supplemented with inorganic Se (57.94%). Hot carcass weight increased (P=0.05) with the increasing of Se levels in the diet. Colour, shear force (SF), cooking and drip loss remained unchanged (P>0.05); however thiobarbituric acid reactive substances was 15.51% higher with inorganic Se compared with organic Se. The selenium concentration in the meat of animals receiving organic selenium was higher (P<0.001) than that of animals receiving sodium selenite, at all levels (0.3; 0.9 and 2.7 mg/kg DM). The meat of animals receiving 2.7 mg of organic Se/kg of DM presented concentration of 372.7 μg Se/kg in the L.dorsi muscle, and the intake of 150 g of this meat by humans provides approximately 100% of the recommended Se intake (55 μg Se/day for adults). Therefore, the use of supranutritional doses of 2.7 mg Se/kg of DM, regardless of source, is a way of naturally producing selenium-enriched meat without compromising performance, carcass characteristics and quality of Nellore bovine meat.     

卫星灵芝液体发酵富集硒锌的研究

研究不同浓度的硒、锌对卫星灵芝菌丝体生长的影响,初步探讨卫星灵芝菌丝体生物富集硒、锌的效应。采用平板培养法及液体发酵法研究锌、硒对卫星灵芝菌丝体生长的影响及富集效应。培养基中不同浓度的亚硒酸钠对菌丝体生长均具有不同程度的抑制作用,但灵芝菌丝体的富硒量随着硒浓度的增加而提高,当亚硒酸钠浓度为40 mg/L时,菌丝体中的生物量、富硒量及富硒转化率最高,分别为1.54%、2 131.55 mg/kg、32.91%;培养基中硫酸锌浓度低于150 mg/L的范围内对卫星灵芝菌丝体生长有明显的促进作用,硫酸锌浓度为60 mg/L时菌丝体中的锌含量和富锌转化率最高,分别为1 142.91 mg/kg、1.76%。培养基中同时添加40 mg/L的亚硒酸钠和60 mg/L硫酸锌,菌丝体生长量1.60%,富硒量301.85 mg/kg,富硒率4.84%;富锌量为540.41 mg/kg,富锌率为5.72%。     

Selenium and Glutathione Peroxidase Levels in Lambs Receiving Feed Supplemented with Sodium Selenite or Selenomethionine

Twenty-one 6 months old female lambs were divided into 7 groups and fed a basal diet containing 0.13 mg Se/kg. The basal diet was further supplemented with 0, 0.1, 0.5 or 1.0 mg Se/kg either as sodium selenite or as selenomethionine, and was fed for 10 weeks. Both feed additives produced an increase in the selenium concentration in the tissues analysed. Significant correlations were found between the concentrations of selenomethionine or sodium selenite added to the feed and the subsequent tissue levels. However, the selenium levels seemed to plateau at approximately 0.5 mg Se/kg of supplemented sodium selenite. The total glutathione peroxidase (GSH-Px) activity of the tissues increased when the selenium supplementation increased from 0 to 0.1 mg/kg for both selenium compounds. With further increase in selenium supplementation the GSH-Px activity in the tissues plateaued except in the blood where the activity continued to rise with increasing selenomethionine supplementation. The selenium dependent GSH-Px activity in the liver rose with increasing selenomethionine supplementation, but approached a plateau when 0.1 mg Se/kg as sodium selenite was added to the feed. The selenium concentration in whole blood responded more rapidly to the selenium supplementation than did GSH-Px activity. The experiment indicates that the optimal selenium concentration in the feed is considerably higher than 0.1 mg Se/kg, and that selenium levels of 1.0 mg/kg in the feed do not result in any risk for the animals or the consumers of the products.     

Effect of the application of selenium on selenium content of soybean and its products

Two experiments were conducted to study the effect of the application of selenium on the selenium content of soybean and its products in two counties with selenium-deficient soil. Selenium-enriched soybean was produced by the application of sodium selenite and Se-enriched fertilizer. The selenium contents of soybeans, soybean protein and okra were determined by hydride-generation atomic fluorescence spectrometry. The results showed that the selenium contents of soybean, soybean protein, and okra were significantly increased by the application of sodium selenite and selenium-enriched fertilizer. Foliar application of selenium provided a higher efficiency for increasing the selenium content of soybean than soil application. Significant differences were found in that soybean cultivars exhibited different accumulation of selenium. There was no remarkable difference in soybean yield, soybean protein, and lipid between selenium and control. The selenium-enriched protein derived from selenium-enriched soybean could be used as a functional ingredient and soybean okra as a selenium-enriched feed for animals for increasing selenium intake.     

富硒酵母和亚硒酸钠对肉仔鸡生长后期日增重影响的Meta分析

富硒酵母和亚硒酸钠对肉仔鸡日增重的影响是否具有差异一直是学界探讨的热点,目前的研究存在着分歧,并因研究众多,实验设计纷繁复杂及传统的综述分析无法定量等原因而不能得出明确的结论,急需尽快判定二者间作用效率的差异．Meta分析是一种统计方法,可将前人具有共同研究目的相互独立的多个研究结果合并、定量分析,得出精确和可靠的结论,避免主观性．本研究中,Meta分析结果表明,在饲料硒浓度0．2mg／kg水平下,亚硒酸钠对肉仔鸡生长后期日增重的作用略大于富硒酵母,但二者无差异显著性（p〉0．05）．     

Bioeffects of selenite on the growth of Spirulina platensis and its biotransformation

The bioeffects of selenium on the growth of Spirulina platensis and the selenium distribution were investigated. S. platensis was batch cultured in Zarrouk medium containing increasing concentrations of sodium selenite. The biotransformation characteristic of selenium was analysed by the determination of the detailed selenium distribution forms. At 35 degrees C, 315.2 microEm(-2) x s(-1), sodium selenite concentrations below 400 mg x l(-1) were found to stimulate algal growth, especially in the range of 0.5-40 mg x l(-1). However, above 500 mg x l(-1) sodium selenite was toxic to this alga with the toxicity being related to the sulfite level in the medium. S. platensis was found to resist higher selenite by reducing toxic Se(IV) to nonsoluble Se(0). Selenium was accumulated efficiently in S. platensis during cultivation with accumulated selenium increasing with selenite concentration in the medium. It was demonstrated that inorganic selenite could be transformed into organic forms through binding with protein, lipids and polysaccharides and other cell components. The organic selenium accounted for 85.1% of the total accumulated selenium and was comprised of 25.2% water-soluble protein-bound, 10.6% lipids-bound and 2.1% polysaccharides-bound selenium. Among the organic fractions lipid possessed the strongest ability to accumulate Se (6.47 mg x kg(-1)). The 14.9% inorganic selenium in S. platensis was composed of Se(IV) (13.7%) and Se(VI) (1.2%).     

Maximizing production of glutathione by amino acid modulation and high-cell-density fed-batch culture of Saccharomyces cerevisiae

In this paper the high-cell-density fed-batch culture and optimal amino acid modulation were combined together to enhance glutathione production in Saccharomyces cerevisiae T65. Ethanol concentration in the broth was an important parameter for feedback control in fed-batch culture. Low ethanol concentration was propitious to both the cell growth and glutathione synthesis. The feedback control of a low ethanol concentration was an efficient way to realize high-cell-density culture and the biomass reached 140 g/L after 57 h fermentation. With optimal amino acid addition to elevate the glutathione content continually, the maximum glutathione yield achieved 2190 mg/L.