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解淀粉芽孢杆菌XT11的黄原胶降解途径的蛋白质组学分析。

Proteomic Analysis of the Xanthan-Degrading Pathway of sp. XT11.

作者信息

Sun Zhen, Liu Huixue, Wang Xueyan, Yang Fan, Li Xianzhen

机构信息

School of Biological Engineering, Dalian Polytechnic University, Dalian 116034, Liaoning, PR China.

出版信息

ACS Omega. 2019 Nov 6;4(21):19096-19105. doi: 10.1021/acsomega.9b02313. eCollection 2019 Nov 19.

Abstract

Xanthan, a highly stable polysaccharide which is not easily degraded by most microorganisms, contains a cellulosic backbone with trisaccharide side chains composed of mannosyl-glucuronyl-mannose attached α-1,3 to alternating glucosyl residues. Different digestion strategies were first applied to demonstrate the complexity about the proteomes of sp. XT11 in xanthan medium and glucose medium. Significantly up-regulated proteins induced by xanthan were screened out by the label-free quantitation of the proteomes of sp. XT11 in xanthan medium and glucose medium. Consequently, 2746 and 2878 proteins were identified in proteomes of sp. XT11 in xanthan medium and glucose medium individually, which represent 80.6 and 84.4% of total protein dataset predicted to be expressed by the gene. In the list of 430 induced proteins containing the proteins specifically expressed or up-regulated in xanthan medium, 19 proteins involved in carbohydrate-active enzymes database and 38 proteins annotated with transporter activity were critical in the degrading pathway of xanthan. Four CAZymes (GH3, GH38, GH9, and PL8) and one ABC transporter (LX1-1GL001097) were verified with quantitative real-time polymerase chain reaction. Four CAZymes (GH3, GH38, GH9, and PL8) were further verified with the enzyme assay. This study suggests a xanthan-degrading pathway in sp. XT11, and other potential xanthan degradation-related proteins still need further investigation.

摘要

黄原胶是一种高度稳定的多糖,不易被大多数微生物降解,它含有一个纤维素主链,其侧链为三糖,由甘露糖基-葡萄糖醛酸基-甘露糖以α-1,3连接到交替的葡萄糖基残基上。首先应用不同的消化策略来证明在黄原胶培养基和葡萄糖培养基中sp. XT11蛋白质组的复杂性。通过对sp. XT11在黄原胶培养基和葡萄糖培养基中的蛋白质组进行无标记定量,筛选出由黄原胶诱导显著上调的蛋白质。结果,在sp. XT11的黄原胶培养基和葡萄糖培养基蛋白质组中分别鉴定出2746和2878种蛋白质,分别占预测由该基因表达的总蛋白质数据集的80.6%和84.4%。在430种诱导蛋白列表中,包括在黄原胶培养基中特异性表达或上调的蛋白质,19种参与碳水化合物活性酶数据库的蛋白质和38种注释有转运活性的蛋白质在黄原胶的降解途径中至关重要。通过定量实时聚合酶链反应验证了四种碳水化合物活性酶(GH3、GH38、GH9和PL8)和一种ABC转运蛋白(LX1-1GL001097)。通过酶活性测定进一步验证了四种碳水化合物活性酶(GH3、GH38、GH9和PL8)。本研究提出了sp. XT11中的黄原胶降解途径,其他潜在的黄原胶降解相关蛋白仍需进一步研究。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f48e/6868878/c414612058b3/ao9b02313_0001.jpg

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