定点突变改变近平滑假丝酵母SCRⅡ催化苯乙酮衍生物底物谱

【目的】通过定点突变技术,改变近平滑假丝酵母短链羰基还原酶Ⅱ(SCRⅡ)催化苯乙酮衍生物的功能,为数种手性芳香醇的生产提供一种高效、安全的新型制备方法。【方法】通过氨基酸序列和蛋白结构比对的方法,选择SCRⅡ的底物结合域中关键氨基酸位点E228实施突变,构建相应的突变株Escherichia coliBL21/pET28a-E228S;以苯乙酮衍生物为底物,对突变株的酶活和生物转化功能进行了分析。【结果】酶活测定结果表明:突变株E.coli BL21/pET28a-E228S催化原始底物2-羟基苯乙酮的酶活仅为原始酶活的25%左右;而催化苯乙酮、4’-甲基苯乙酮、4’-氯苯乙酮的酶活是突变前的7-20倍。突变株E.coli BL21/pET28a-E228S生物转化2-羟基苯乙酮,获得产物(S)-苯基乙二醇的得率不超过10%,而以苯乙酮、4’-甲基苯乙酮、4’-氯苯乙酮为底物时,生物转化产物光学纯度维持在99%,得率高达80%以上。【结论】对底物结合域中的关键氨基酸实施突变,提高了SCRⅡ催化苯乙酮衍生物的底物广谱性,拓展了该酶的生物功能,为理性改造短链羰基还原酶的不对称还原催化功能和手性芳香醇的制备提供了新型途径。     

微生物协同Fe0氧化的环境修复作用研究进展

零价铁(Fe0)具有高效还原转化多种污染物的能力,但不能实现污染物的矿化作用。微生物与Fe0的协同作用过程,以微生物为主导,Fe0起促进作用,可有效提高多种污染物的降解效率,实现污染物的彻底脱毒与无害化,因此利用微生物协同Fe0氧化进行环境修复具有广阔的应用前景。本文从微生物协同Fe0氧化的作用机理、菌种多样性及其在环境修复中的应用等研究进展进行综述,提出微生物协同Fe0氧化的环境修复研究中存在的主要问题和重点研究方向,以期在更全面、深入地认识这一过程的基础上,充分发挥其在环境修复中的作用。     

Comparative Study of Carbon Storage in Different Forest Stands in Subtropical China

Selection and development of tree species with high fixing CO₂ capacity is an increasing problem worldwide. A comparative study on carbon fixation ability of three forest stands was conducted at Linlong Mountain, Li’nan County, Zhejiang Province, China. The results showed that total carbon storage in the ecosystems of Moso bamboo, Chinese fir, and Masson pine stands were 104.83, 95.66, and 96.49 t C/ha, respectively. The spatial distribution of carbon storage in the three ecosystems decreased in the order: soil > tree story > the vegetation under the forests. Carbon storage in the soils under Moso bamboo, Chinese fir, and Masson pine stands accounted for 65.3, 61.4, and 55.6% of the total CSs, respectively. The Moso bamboo forest ecosystem fixed 1.69 and 1.63 times as much C (9.64 t C/ha/year) as the Chinese fir and Masson pine forest ecosystems, respectively.     

A genetic map of Xenopus tropicalis

We present a genetic map for Xenopus tropicalis, consisting of 2886 Simple Sequence Length Polymorphism (SSLP) markers. Using a bioinformatics-based strategy, we identified unique SSLPs within the X. tropicalis genome. Scaffolds from X. tropicalis genome assembly 2.0 (JGI) were scanned for Simple Sequence Repeats (SSRs); unique SSRs were then tested for amplification and polymorphisms using DNA from inbred Nigerian and Ivory Coast individuals. Thus identified, the SSLPs were genotyped against a mapping cross panel of DNA samples from 190 F2 individuals. Nearly 4000 SSLPs were genotyped, yielding a 2886-marker genetic map consisting of 10 major linkage groups between 73 and 132 cM in length, and 4 smaller linkage groups between 7 and 40 cM. The total effective size of the map is 1658 cM, and the average intermarker distance for each linkage group ranged from 0.27 to 0.75 cM. Fluorescence In Situ Hybridization (FISH) was carried out using probes for genes located on mapped scaffolds to assign linkage groups to chromosomes. Comparisons of this map with the X. tropicalis genome Assembly 4.1 (JGI) indicate that the map provides representation of a minimum of 66% of the X. tropicalis genome, incorporating 758 of the approximately 1300 scaffolds over 100,000 bp. The genetic map and SSLP marker database constitute an essential resource for genetic and genomic analyses in X. tropicalis.