Gene Cloning,Expression and Characterization of a Novel Xylanase from the Marine Bacterium,Glaciecola mesophila KMM241

Marine xylanases are rather less studied compared to terrestrial xylanases. In this study, a new xylanase gene, xynB, was cloned from the marine bacterium, Glaciecola mesophila KMM241, and expressed in Escherichia coli. xynB encodes a multi-domain xylanase XynB of glycoside hydrolase (GH) family 8. The recombinant XynB comprises an N-terminal domain (NTD) with unknown function and a catalytic domain, which is structurally novel among the characterized xylanases of GH family 8. XynB has the highest identity (38%) to rXyn8 among the characterized xylanases. The recombinant XynB showed maximal activity at pH 6–7 and 35 °C. It is thermolabile and salt-tolerant. XynB is an endo-xylanase that demands at least five sugar moieties for effective cleavage and to hydrolyze xylohexaose and xylopentaose into xylotetraose, xylotriose and xylobiose. NTD was expressed in Escherichia coli to analyze its function. The recombinant NTD exhibited a high binding ability to insoluble xylan and avicel and little binding ability to chitosan and chitin. Since the NTD shows no obvious homology to any known carbohydrate-binding module (CBM) sequence in public databases, XynB may contain a new type of CBM.     

复合微生物菌肥KMM减缓苹果再植病害机制

为揭示木美土里复合微生物菌肥（Kimidori microbial manure,KMM）促进再植苹果树的生长及减缓苹果再植病的作用机制,采用土壤常规农业化学分析法和Illumma MiSeq高通量测序技术比较分析KMM和常规有机肥（对照）处理后再植苹果树的生长指标、理化因子、根际土壤酶活性以及细菌群落结构和功能的变化。结果表明,施用KMM后,果树的株高增长率、干径增长率、叶绿素含量、分枝个数和分枝长度等生长指标均显著升高,有机质含量显著增加,中性磷酸酶、脲酶、蔗糖酶和过氧化氢酶活性均显著提高,同时增加了再植土壤细菌群落的丰富度和多样性。从采集土壤中分离得到51株细菌并进行盆栽试验,其中经KMM-15、KMM-37和KMM-50菌株处理后海棠苗叶绿素含量、株高、茎粗均显著提高,经鉴定这3株菌株为贝莱斯芽胞杆菌Bacillus velezensis。表明春秋2次施用KMM提高了土壤有机质含量和酶活性,并重塑了根际土壤中细菌群落结构,进而缓解苹果再植病害发生程度,促进果树生长。