一株产酶真菌Phaeophlebiopsis sp.转录组分析

对从药渣中分离的真菌ZYJHYZ254进行鉴定及产酶活性研究,从转录组分析菌株不同生长时期差异表达基因对其生长发育及产酶调控的影响,以筛选高产木质纤维素水解酶真菌,寻找调控关键基因。鉴定ZYJHYZ254为拟暗射脉菌Phaeophlebiopsis sp.,产酶在第5-7天最高。从生长3 d与7 d的菌丝中共检测到1 232个差异基因,以3 d的菌丝为对照,显著上调、下调基因分别有826及406个,基因注释和GO、KEGG功能富集分析结果表明差异表达基因主要与蛋白质合成、代谢及酶合成相关。此外,共有387个CAZymes基因表达,GH数量最多,约占49.61%,其次为AA (97)与GT (62),约占25.06%与16.02%。GH16 (24个)占GH的12.50%,含量最多,主要编码葡萄糖苷酶、木聚糖酶等,AA中AA3 (37个)占比38.14%,编码氧化酶、脱氢酶等。结果表明ZYJHYZ254中生长3 d与7 d的菌丝经功能富集分析发现差异表达基因主要与蛋白质合成、代谢,以及酶合成相关。进一步研究发现在两个生长时期中CAZymes基因表达最多的是GH16与AA3,预示了该菌葡萄糖苷酶、木聚糖酶、β-半乳糖苷酶、氧化酶与脱氢酶含量最丰富,对降解特殊生物质中的木质纤维素具有重要意义。     

合成生物制造2022

本文对2022年《生物工程学报》发表的与合成生物制造相关的综述和研究论文进行了评述,重点讨论了DNA测序、DNA合成、DNA编辑、基因表达调控和数学细胞模型等底层技术,酶的设计、改造和应用技术,化学品生物催化、氨基酸及其衍生物、有机酸、天然化合物、抗生素与活性肽、功能多糖、功能蛋白质等重要产品的生物制造技术,一碳化合物和生物质原料利用技术以及合成微生物组技术,以帮助读者从一个侧面了解合成生物制造相关技术和产业的发展情况。     

Photosynthetic characteristics and effect of ATP in transgenic rice with phospho<Emphasis Type="Italic">enol</Emphasis>pyruvate carboxylase and pyruvate orthophosphate dikinase genes

In the untransformed rice (WT) and transgenic rice with the PEPC and PPDK genes (CK) we determined activities of C₄ photosynthetic enzymes, photosynthetic response to irradiance and temperature, the metabolic index of active oxygen, and the yield component factors. The activities of C₄ photosynthetic enzymes in WT were very low, while those of corresponding enzymes in CK were highly observable. Moreover, after adenosine triphosphate (ATP) treatment, and under high irradiance and high temperature, the net photosynthetic rate of CK increased by 17 and 12 %, respectively, as compared to that achieved without ATP treatment. The resistance of CK against photo-oxidation was enhanced under these conditions, and CK yield increased by 15 %. ATP treatment enhanced the photosynthetic productivity of CK, thereby proving that ATP is the key factor in enhancing the photosynthetic capacity of transgenic rice with C₄ gene. Our new technical approach can be used in breeding rice with high photosynthetic efficiency and high grain yield.     

Alterations of Antioxidant Enzymes and Oxidative Damage to Macromolecules in Different Organs of Rats During Aging

Oxygen free radicals have been hypothesized to play an important role in the aging process. To investigate the correlation between the oxidative stress and aging, we have determined the levels of oxidative protein damage and lipid peroxidation in the brain and liver, and activities of antioxidant enzymes in the brain, liver, heart, kidney, and serum from the Fisher 344 rats at ages of 1, 6, 12, 18, and 24 months. The results showed that the level of oxidative protein damage (measured as carbonyl content) in the brain and liver was significantly higher in older animals than in young animals. No statistical difference was observed in the lipid peroxidation of the liver and brain between young and old animals. The activities of antioxidant enzymes in most tissues displayed an age-dependent decline. Superoxide dismutases in the heart, kidney, and serum, glutathione peroxidase activities in the serum and kidney, and catalase activities in the brain, liver, and kidney, significantly decreased during aging. Cytochrome c oxidase, an enzyme involved in electron transport in mitochondria, initially increased, but subsequently decreased in the aged brain, whereas no significant alteration was observed in the liver mitochondrial antioxidant enzymes. The present studies suggest that the accumulation of oxidized proteins during aging is most likely to be linked with an age-related decline of antioxidant enzyme activities, whereas lipid peroxidation is less sensitive to predict the aging process.     

Polyelectrolyte-coated microcapsules and their potential applications to biotechnology

Polyelectrolyte-coated microcapsules can be prepared by adsorption of polyions onto microcapsule surfaces in aqueous solutions under appropriate pH and ionic conditions. The resulting polyelectrolyte-coated microcapsules provide a promising tool for studying pH-induced configurational changes in polyions adsorbed onto hydrophobic membranes (capsule walls). An interesting application of polyelectrolyte-coated microcapsules is the pH-sensitive on/off control of microencapsulated enzyme reactions through alterations in the substrate permeability of the capsule wall by pH-conditioned configurational changes in the adsorbed polyion layer. This paper presents an overview of pH-induced conformational changes of polyelectrolytes in solutions, preparation of polyelectrolyte-coated microcapsules with an immobilized enzyme, and on/off control of the respective enzyme reactions by pH adjustment. This revised version was published online in July 2006 with corrections to the Cover Date.     

斑蝥素对粘虫几种代谢酶及多酚氧化酶的影响

为进一步探讨斑蝥素的杀虫作用机理, 本研究采用叶碟饲喂法处理粘虫Mythimna separata（Walker）5龄幼虫, 测定了饲喂处理后6, 12, 24, 36和48 h试虫体内羧酸酯酶(CarE)、酸性磷酸酯酶(ACP)、碱性磷酸酯酶(ALP)、谷胱甘肽 S-转移酶(GST)和细胞色素P450 Ο-脱甲基酶及多酚氧化酶(PPO)的活性。结果表明: 斑蝥素可显著激活羧酸酯酶, 处理后48 h酶比活力最大, 为同期对照的1.68倍; 酸性磷酸酯酶在处理后6和12 h活性变化不明显, 处理24 h后逐渐被激活, 且与对照差异显著(P＜0.05); 明显抑制碱性磷酸酯酶, 且随着处理时间的延长, 抑制作用增强; 对Ο-脱甲基酶表现为先抑制后激活的趋势; 谷胱甘肽S-转移酶表现出先激活后抑制的影响; 离体活体条件下均可显著抑制PPO活性。可见, 斑蝥素可明显影响昆虫的代谢酶系, 且其对碱性磷酸酶和多酚氧化酶的抑制作用可能与其毒杀作用有关。     

‘赛蜜酥1号’枣及其芽变品系果实的性状差异北大核心CSCD

该试验以‘赛蜜酥1号’枣及其芽变品系‘赛蜜酥2号’果实为试验材料,运用流式细胞术对它们的倍性进行鉴定,采用石蜡切片法进行果实细胞组学的观察,并对两者果实生长发育过程中的内外观品质及决定果实口感的蔗糖代谢相关酶活性进行比较分析,为深入研究芽变对枣果实口感、品质造成的影响提供理论支持。结果表明:(1)‘赛蜜酥2号’芽变品系的细胞倍性未发生改变,仍为二倍体。(2)两个枣品种(系)果实在外部形态上有明显区别,‘赛蜜酥1号’为卵圆型,‘赛蜜酥2号’为扁圆型,且后者的果形指数成熟时大于1,单果质量高于前者,核小、可食率更高,皮薄果实更加酥脆;‘赛蜜酥1号’果皮的蜡质层厚度一直大于其芽变枣,两者角质层厚度和表皮层厚度的变化趋势基本一致,但厚度之间具有显著差异。(3)影响果实细胞生长、分裂,果实脱落的各类激素水平在品种间均存在显著差异,‘赛蜜酥2号’IAA和GA_(3)含量显著高于‘赛蜜酥1号’,使之果形更大。(4)两个枣品种(系)果实可溶性糖、可滴定酸、维生素C、植物淀粉、可溶性蛋白含量等随发育时期的变化趋势一致,但含量水平之间存在差异,‘赛蜜酥2号’果实的糖含量、维生素C含量更高,可滴定酸含量更低,口感更甘甜。(5)两品种枣果实的蔗糖代谢相关酶活性在果实迅速生长的膨大期都存在着显著差异,也导致‘赛蜜酥2号’果实甜度明显高于‘赛蜜酥1号’。研究发现,两个枣品种均为二倍体,它们在果实外部形态上存在明显区别,易于分辨;芽变品系‘赛蜜酥2号’果实更大,核小、可食率更高,皮薄更加酥脆,口感更甘甜。     

生物酶/γ-Al2O3小球催化氧化柴油脱硫性能研究

通过吸附法将生物酶负载在γ-Al₂O₃小球载体上,并对生物酶/γ-Al₂O₃及载体进行扫描电镜(SEM)、比表面积分析(BET)、傅里叶红外光谱(FT-IR)及圆二色谱(CD)表征。结果表明:生物酶被吸附在载体上。将制备的生物酶/γ-Al₂O₃催化真实柴油氧化脱硫,考察了反应温度、反应流速和酶溶液浓度对真实柴油脱硫效果的影响,并对脱硫效果进行定性及定量分析;进一步对脱硫工艺条件进行响应面设计优化,找出最优反应条件。实验结果显示:反应温度49℃、反应流速1.0 mL/min、酶溶液浓度15.5%(酶载量为28.13 g),得出的最优脱硫率为93.16%;最后考察了该固定化酶的重复使用性能,该催化剂使用7次活性无明显降低,表明该固定化酶催化氧化柴油脱硫效果显著,具有潜在的工艺应用价值。     

Mitochondrial DNA as a genetic marker for brown trout, Salmo trutta L., populations

Mitochondrial DNA was examined in natural and hatchery-reared stocks of brown trout, using different methods of restriction analysis. The methods included the development of a brown trout mt DNA hybridization probe through cloning part of the brown trout mitochondrial genome. In addition, fragments were analysed by ethidium bromide staining and end-labelling. The relative merits of each of these methods in assessing levels of genetic relatedness between the natural and hatchery-reared brown trout stocks were evaluated. In addition, the study revealed a diagnostic mtDNA restriction pattern which could be used as a genetic marker for the discrimination of these two groups of brown trout.