本文選題：微紫青霉　切入點(diǎn)：酸性木聚糖酶　出處：《食品科學(xué)》2017年14期 　論文類型：期刊論文

【摘要】：基于酸性木聚糖酶在飼料及釀酒行業(yè)良好的應(yīng)用前景,通過(guò)基因組步移的方法克隆得到微紫青霉酸性木聚糖酶的全長(zhǎng)基因xynA,然后采取重疊延伸PCR技術(shù)進(jìn)行內(nèi)含子的切除獲得xynA的cDNA序列,并對(duì)其進(jìn)行了生物信息學(xué)分析。序列分析結(jié)果顯示xynA基因全長(zhǎng)720 bp,內(nèi)含子63 bp,cDNA全長(zhǎng)657 bp。推測(cè)該木聚糖酶編碼信號(hào)肽28個(gè)氨基酸,成熟肽190個(gè)氨基酸;預(yù)測(cè)該蛋白為分子質(zhì)量20.61 kD、等電點(diǎn)7.0的親水性蛋白,且分子內(nèi)不含二硫鍵。與其他真菌來(lái)源的GH11族耐酸性木聚糖酶進(jìn)行序列比對(duì),結(jié)果顯示該酶的相應(yīng)位置具有特征天冬氨酸殘基Asp,且具有糖苷水解酶11族的保守區(qū)域特征以及典型的"右手半握"狀結(jié)構(gòu),重組木聚糖酶基因xynA能夠在大腸桿菌中成功表達(dá),比酶活力達(dá)220.5 U/mg。
[Abstract]:Based on the good application prospect of acid xylanase in feed and wine industry, The full-length gene XynAof acid xylanase of Penicillium microphylla was cloned by genomic step, and the cDNA sequence of xynA was obtained by excision of intron by overlapping extension PCR. The sequence analysis showed that the full length of xynA gene was 720bpand the full length of intron 63bp cDNA was 657bp. it was deduced that the xylanase encode signal peptide of 28 amino acids and mature peptide of 190 amino acids. The protein was predicted to be a hydrophilic protein with a molecular weight of 20.61 kD and an isoelectric point of 7.0, and no disulfide bond was found in the molecule. The protein was sequenced to GH11 group acidtolerant xylanase from other fungi. The results showed that the enzyme had the characteristic aspartic acid residue AspS, the conserved region of the glycoside hydrolase group 11 and the typical right-hand half-grip structure. The recombinant xylanase gene xynA could be successfully expressed in Escherichia coli. The specific enzyme activity was 220.5Umg.
【作者單位】： 北京工商大學(xué)北京食品營(yíng)養(yǎng)與人類健康高精尖創(chuàng)新中心;北京工商大學(xué)北京市食品添加劑工程技術(shù)研究中心;北京工商大學(xué)食品學(xué)院;
【基金】：國(guó)家自然科學(xué)基金面上項(xiàng)目(31371723)
【分類號(hào)】：Q78

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