The role of peroxisomes in xylose alcoholic fermentation in the engineered Saccharomyces cerevisiae

Xylose is a second‐most abounded sugar after glucose in lignocellulosic hydrolysates and should be efficiently fermented for economically viable second‐generation ethanol production. Despite significant progress in metabolic and evolutionary engineering, xylose fermentation rate of recombinant Saccharomyces cerevisiae remains lower than that for glucose. Our recent study demonstrated that peroxisome‐deficient cells of yeast Ogataea polymorpha showed a decrease in ethanol production from xylose. In this work, we have studied the role of peroxisomes in xylose alcoholic fermentation in the engineered xylose‐utilizing strain of S. cerevisiae. It was shown that peroxisome‐less pex3Δ mutant possessed 1.5‐fold decrease of ethanol production from xylose. We hypothesized that peroxisomal catalase Cta1 may have importance for hydrogen peroxide, the important component of reactive oxygen species, detoxification during xylose alcoholic fermentation. It was clearly shown that CTA1 deletion impaired ethanol production from xylose. It was found that enhancing the peroxisome population by modulation the peroxisomal biogenesis by overexpression of PEX34 activates xylose alcoholic fermentation.     

蛹虫草菌生物合成虫草素的研究进展

蛹虫草Cordyceps militaris是我国传统的药用真菌,虫草素是蛹虫草的主要活性成分,具有抗癌、抗肿瘤、抗病毒等多种生理功能。蛹虫草菌液体发酵是最有希望实现高效生产虫草素的途径,但现阶段生产强度低,亟需应用发酵工程及代谢工程手段提高虫草素产量。文中对液体发酵体系中培养基组分(碳/氮源、前体物质、金属离子等)和培养条件(pH、溶氧量、光照等)对虫草素产量的影响进行了总结,并对虫草素的分离纯化、生物合成基因簇及合成代谢途径进行了阐述,最后探讨了实现虫草素高效生产的关键环节。     

一株近玫色锁掷酵母的分离鉴定及其产胞外乳糖酶性质的研究

从广州徐闻农垦丰收农场土壤样品中分离到一株产乳糖酶菌株,结合菌株形态特征和ITS基因序列同源性分析结果,表明该菌株为Saccharomycetes sp.的未定种,其系统分类学关系与近玫色锁掷酵母Sporidiobolus pararoseus strain AUMC 7791(JQ425362.1)最近,故命名为:近玫色锁掷酵母XWSP1(Sporidiobolus pararoseus XWSP1),该菌种保藏号为CCTCC NO:M2019119。该菌发酵液能水解邻硝基酚-β-D-半乳糖苷生成黄色的邻硝基酚,具有产乳糖酶性能。优化了该菌株发酵培养条件,结果表明:该菌株在蛋白胨15 g/L、酵母粉20 g/L、半乳糖20 g/L和初始pH 7.0的培养基中,以1×106 CFU/mL接种浓度、4%接种比例,28℃180 r/min恒温振荡培养54 h时,菌株分泌的胞外乳糖酶具有最高的活力。酶学性质初步研究结果表明,该胞外乳糖酶在pH 7.0的反应条件下酶活力最高,Ca^2+、Mn^2+、Mg^2+和Cu^2+对酶活有不同程度抑制作用,其中Cu^2+对酶活抑制作用最强。     

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。     

厌氧发酵液中溶解性有机物对活性污泥合成聚羟基脂肪酸酯影响的研究进展

利用活性污泥微生物将剩余污泥发酵液中的挥发性脂肪酸(Volatile fatty acids,VFAs)转化为聚羟基脂肪酸酯(Polyhydroxyalkanoates,PHA)是目前环境生物技术领域的研究热点。但针对发酵液中非VFAs物质(主要是溶解性有机物,Dissolved organic matter,DOM)对活性污泥合成PHA的影响目前仍未有统一的结论,需要进行较全面的分析和总结。因此,文中首先介绍了剩余污泥发酵液中DOM的主要特性及常见分析方法,然后从微生物学、代谢调控、污泥性质等角度分析了DOM中几种主要组分(多聚糖、蛋白类物质、腐殖质)对活性污泥合成PHA的影响。综合各研究结果表明高浓度DOM不利于PHA的生产,但适量DOM的存在有利于污泥性质的稳定,同时能降低PHA提纯成本。最后提出了相应的策略来实现对活性污泥利用DOM合成PHA的调控,以期为高效利用厌氧发酵液中DOM实现PHA合成提供解决思路。     

短链硫酯酶缺失对大肠杆菌合成聚羟基丁酸乳酸酯的影响

聚羟基脂肪酸酯 (Polyhydroxyalkanoates,PHAs) 是一种具有优质生物相容性的可降解生物基材料,其理化性质优越,具备替代石油基塑料的潜力。P(3HB-co-LA) 是PHAs的一种,融合了聚乳酸 (Polylactic acid,PLA) 和聚3-羟基丁酸 (poly(3-hydroxybutyrate),P(3HB)) 共同的优点,具有更好的韧性和透明度。文中首先在大肠杆菌MG1655中通过质粒表达外源基因phaA、phaB、phaCm和pctth,发酵生产出乳酸含量为23.8 mol%的P(3HB-co-LA),在此基础上缺失dld基因得到菌株WXJ01-03,其合成的聚合物中乳酸组分含量提升至37.2 mol%。当硫酯酶基因ydiI和yciA基因继续被敲除后,生产的共聚物中乳酸组分进一步提升至42.3 mol%和41.1 mol%。最后将3个基因dld、yciA和ydiI同时缺失得到重组菌株WXJ03-03,并通过该重组菌株获得了乳酸组分含量为46.1 mol%的共聚物。通过比较不同碳源的发酵结果得知,木糖有利于提高共聚物中乳酸组分含量。上述实验结果表明,在木糖发酵中短链硫酯酶基因缺失阻碍了大肠杆菌胞内的LA-CoA被降解,可有效提高聚合物中乳酸组分的摩尔百分比。     

维生素C生物合成相关脱氢酶研究进展

维生素C是一种人体必需的维生素,在食品制药等领域拥有巨大的市场。工业上维生素C主要以微生物发酵生产的2-酮基-L-古龙酸为前体,然后通过内酯化反应获得。微生物发酵中,山梨糖途径和葡萄糖酸途径因为转化率高一直是研究的热点。文中从维生素C生物合成相关脱氢酶的角度阐述了:山梨糖途径和葡萄糖酸途径中关键脱氢酶在定位、底物谱、辅因子和电子传递上的特点;山梨糖途径和葡萄糖酸途径中面临的主要问题和改造策略等。最后讨论了维生素C生物合成中山梨糖途径和葡萄糖酸途径可能的研究方向。     

玉米雄穗性状遗传结构与形成分子机制*

玉米为雌雄同株异花植物,其雄穗着生于植株顶部,雌穗腋生。雄穗一方面需产生足量花粉以保证雌穗授粉结实,另一方面由于对下部叶片的遮蔽作用和自身营养需求,其生长发育会同时影响叶片光合作用效率和能量分配,因此优化雄穗结构是提高玉米产量的重要措施之一。玉米雄穗性状包括雄穗分枝数、雄穗分枝长度、雄穗主轴长度、雄穗分枝总长度、雄穗分枝角度等,均为多基因控制的数量性状。自20世纪90年代,研究者开始利用数量性状位点(quantitative trait locus,QTL)定位方法解析玉米雄穗性状遗传结构;随着玉米自交系B73等参考基因组释放,以及DNA微阵列、基因组重测序等高通量基因分型技术的日益成熟,全基因组关联分析(genome-wide association study, GWAS)成为数量性状遗传研究的主流方法,目前已鉴定出大量玉米雄穗性状遗传位点。通过总结雄穗性状遗传定位研究结果,构建一致性图谱并挖掘定位热点区间,有助于进一步了解雄穗性状遗传结构特征及指导雄穗性状候选基因克隆。此外,通过对调控雄穗发育的已知基因进行功能分类,可为解析玉米雄穗发育的遗传网络和调控通路提供理论支撑。     

Effects of faba bean,blue lupin and rapeseed meal supplementation on nitrogen digestion and utilization of dairy cows fed grass silage-based diets

There is increasing interest in using locally produced protein supplements in dairy cow feeding. The objective of this experiment was to compare rapeseed meal (RSM), faba beans (FBs) and blue lupin seeds (BL) at isonitrogenous amounts as supplements of grass silage and cereal based diets. A control diet (CON) without protein supplement was included in the experiment. Four lactating Nordic Red cows were used in a 4 × 4 Latin Square design with four 21 d periods. The milk production increased with protein supplementation but when expressed as energy corrected milk, the response disappeared due to substantially higher milk fat concentration with CON compared to protein supplemented diets. Milk protein output increased by 8.5, 4.4 and 2.7% when RSM, FB and BL were compared to CON. The main changes in rumen fermentation were the higher propionate and lower butyrate proportion of total rumen volatile fatty acids when the protein supplemented diets were compared to CON. Protein supplementation also clearly increased the ruminal ammonia N concentration. Protein supplementation improved diet organic matter and NDF digestibility but efficiency of microbial protein synthesis per kg organic matter truly digested was not affected. Flow of microbial N was greater when FB compared to BL was fed. All protein supplements decreased the efficiency of nitrogen use in milk production. The marginal efficiency (amount of additional feed protein captured in milk protein) was 0.110, 0.062 and 0.045 for RSM, FB and BL, respectively. The current study supports the evidence that RSM is a good protein supplement for dairy cows, and this effect was at least partly mediated by the lower rumen degradability of RSM protein compared to FB and BL. The relatively small production responses to protein supplementation with simultaneous decrease in nitrogen use efficiency in milk production suggest that economic and environmental consequences of protein feeding need to be carefully considered.