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81.
82.
Xiaoyan Guo Yunfang Gao Fangzheng Liu Yong Tao Haibo Jin Jianjun Wang Sheng Wu 《Microbial biotechnology》2023,16(6):1333-1343
R-1,3-butanediol (R-1,3-BDO) is an important chiral intermediate of penem and carbapenem synthesis. Among the different synthesis methods to obtain pure enantiomer R-1,3-BDO, oxidation–reduction cascades catalysed by enzymes are promising strategies for its production. Dehydrogenases have been used for the reduction step, but the enantio-selectivity is not high enough for further organic synthesis efforts. Here, a short-chain carbonyl reductase (LnRCR) was evaluated for the reduction step and developed via protein engineering. After docking result analysis with the substrate 4-hydroxy-2-butanone (4H2B), residues were selected for virtual mutagenesis, their substrate-binding energies were compared, and four sites were selected for saturation mutagenesis. High-throughput screening helped identify a Ser154Lys mutant which increased the catalytic efficiency by 115% compared to the parent enzyme. Computer-aided simulations indicated that after single residue replacement, movements in two flexible areas (VTDPAF and SVGFANK) facilitated the volumetric compression of the 4H2B-binding pocket. The number of hydrogen bonds between the stabilized 4H2B-binding pocket of the mutant enzyme and substrate was higher (from four to six) than the wild-type enzyme, while the substrate-binding energy was decreased (from −17.0 kJ/mol to −29.1 kJ/mol). Consequently, the catalytic efficiency increased by approximately 115% and enantio-selectivity increased from 95% to 99%. Our findings indicate that compact and stable substrate-binding pockets are critical for enzyme catalysis. Lastly, the utilization of a microbe expressing the Ser154Lys mutant enzyme was proven to be a robust process to conduct the oxidation–reduction cascade at larger scales. 相似文献
83.
84.
Lu‐Xian Liu Pan Deng Meng‐Zhen Chen Li‐Min Yu Joongku Lee Wei‐Mei Jiang Cheng‐Xin Fu Fu‐De Shang Pan Li 《植物分类学报:英文版》2023,61(1):99-114
Oresitrophe and Mukdenia (Saxifragaceae) are epilithic sister genera used in traditional Chinese medicine. The taxonomy of Mukdenia, especially of M. acanthifolia, has been controversial. To address this, we produced plastid and mitochondrial data using genome skimming for Mukdenia acanthifolia and Mukdenia rossii, including three individuals of each species. We assembled complete plastomes, mitochondrial CDS and nuclear ribosomal ETS/ITS sequences using these data. Comparative analysis shows that the plastomes of Mukdenia and Oresitrophe are relatively conservative in terms of genome size, structure, gene content, RNA editing sites and codon usage. Five plastid regions that represent hotspots of change (trnH-psbA, psbC-trnS, trnM-atpE, petA-psbJ and ccsA-ndhD) are identified within Mukdenia, and six regions (trnH-psbA, petN-psbM, trnM-atpE, rps16-trnQ, ycf1 and ndhF) contain a higher number of species-specific parsimony-informative sites that may serve as potential DNA barcodes for species identification. To infer phylogenetic relationships between Mukdenia and Oresitrophe, we combined our data with published data based on three different datasets. The monophyly of each species (Oresitrophe rupifraga, M. acanthifolia and M. rossii) and the inferred topology ((M. rossii, M. acanthifolia), O. rupifraga) are well supported in trees reconstructed using the complete plastome sequences, but M. acanthifolia and M. rossii did not form a separate clade in the trees based on ETS + ITS data, while the mitochondrial CDS trees are not well-resolved. We found low recovery of genes in the Angiosperms353 target enrichment panel from our unenriched genome skimming data. Hybridization or incomplete lineage sorting may be the cause of discordance between trees reconstructed from organellar and nuclear data. Considering its morphological distinctiveness and our molecular phylogenetic results, we strongly recommend that M. acanthifolia be treated as a distinct species. 相似文献
85.
一套新的籼稻初级三体的选育和细胞学鉴定 总被引:26,自引:0,他引:26
以秋水仙素处理中籼3037幼苗,从其后代中获得同源三倍体1株。为选育成套初级三体,将中籼3037同源三倍体茎节幼芽置于试管中离体繁殖,繁殖后的同源三倍体种于田间,授以正常中籼3037的花粉。获得3013粒种子,成苗1063株。通过染色体数目检查,共获得302株三体植株。这些三体按其形态特征可分为12类,对每类三体植株分别通过粗线期的染色体鉴定,获得了12种初级三体,依据额外染色体长度递减的顺序分别定名为三体1、三体2、………三体12。 相似文献
86.
红原鸡与家鸡的亲缘关系研究 总被引:15,自引:2,他引:13
对中国红原鸡滇地亚种和海南亚种与我国茶花鸡,泰和鸡和寿光鸡等地方鸡种以及芦花鸡,洛岛红等外国鸡种进行了血型(3个位点,13个等位基因),蛋白质(酶)多态(5个位点,11个等位基因)和DNA指纹分析,结果表明,红原鸡与茶花鸡(原始型品种)的亲缘关系较近;与泰和鸡,寿光鸡,芦花鸡,洛岛红(进化型品种)的亲缘关系较远,呈红原鸡-茶花鸡-泰和鸡,寿光鸡或芦花鸡,洛岛红这样一个进化阶梯,以上结果与国外资料( 相似文献
87.
脏器微血管对荧光素钠通透性的实验方法 总被引:4,自引:1,他引:3
大鼠颈动脉注射1%FlNa,荧光显微镜下活体观察肠系膜微血管血流状态及FlNa的渗出情况,并在不同时间点经股动脉采血,测定血浆内FlNa浓度随时间的变化,利用组织匀浆测定不同脏器中FlNa的分布,再辅以冰冻切片进行观察。活体观察发现,FlNa注入体内后,经微血管迅速向周围组织渗出,最后汇集于淋巴管,血浆及组织匀浆FlNa浓度的测定表明,FlNa浓度随时间的变化呈指数衰减,各脏器FlNa的分布极不相同。冰冻切片也显示了同样的分布差别。这些结果表明,我们所建立的方法可直观、定量地反映FlNa在微血管的通透情况。 相似文献
88.
89.
关中小麦品种产量构成因素的相互关系和产量育种目标 总被引:7,自引:0,他引:7
对关中地区不同年代及当前大面积推广的小麦品种(系)的产量及其构成因素进行了遗传分析和通径分析。结果表明:陕西关中地区近期小麦品种的产量育种目标的趋势是在提高现有穗粒数的基础上,保持现有的穗数(700万/hm^2)及千粒重(38g左右),以求在提高品种产量潜力的同时,保持或增加品种的稳定性能;或是大幅度提高穗粒数(35粒左右),适当提高千粒重(40g左右),适当降低穗数(600万/hm^2左右),以 相似文献
90.
高压静电场处理沙棘插条生根状况的初步研究 总被引:2,自引:0,他引:2
用高压静电场处理沙棘插条研究生根状况,方法简便易于处理,该装置适于进行大规模的处理,从目前研究表明:高压静电场对沙棘插条有正刺激效应,并以电场强度为3.9kv/cm,时间为40分钟处理最佳。 相似文献