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首页> 外文会议>International conference on applied biotechnology >Enhancement of Gellan Production in Sphingomonas paucimobilis JLJ by Heterogeneous Expression of Vitreoscilla Hemoglobin
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Enhancement of Gellan Production in Sphingomonas paucimobilis JLJ by Heterogeneous Expression of Vitreoscilla Hemoglobin

机译：玻璃体血球蛋白血红蛋白的异源表达增强了鞘氨醇单胞菌JLJ的结冷蛋白产量

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A heterologous expression of Vitreoscilla hemoglobin (VHb) for improving cell growth and fermentation products has been successfully demonstrated in Various hosts. To improve a commercially used strain for gellan, vgb gene was expressed in Sphingomonas paucimobilis JLJ under the control of the oxygen-dependent vgb gene promoter (P_(vgb)). The promoter was maximally induced under microaerobic conditions. Biochemical activity of expressed vgb was confirmed by PCR, SDS-PAGE, and the CO-difference spectra analysis which exhibited a characteristic absorption maximum near 420 nm. The results showed that the expression of vgb could enhance cell growth. The composition of fermentation medium and the physical conditions of cultivations were investigated. The optimum medium consists of (g/L): Sucrose 30, Soybean meal 5, NaNC_3 1, KH_2PO_4 1, K_2HPO_4 1.5, MgSO_4 0.6. The optimum fermentation conditions were confirmed: pH 6.0, seed age 16 h, inoculum volume 10 %, 200 rpm, 30 °C, and the maximum gellan yield of vgb~+ recombinant was 1.713 % +- 0.02 % after cultivated 72 h, 21 % higher than that of the parental strain. VHb exhibited positive effect on cell growth and optimize gellan yield. The results provided potential applications of vgb gene in other industrially important strains to solve the increasing demand of aerobic processes in polysaccharides production.
机译：玻璃体血红蛋白（VHb）用于改善细胞生长和发酵产物的异源表达已在各种宿主中成功证明。为了改进商购的结冷胶菌株，在氧依赖性vgb基因启动子（P_（vgb））的控制下，在鞘脂单胞菌JLJ中表达了vgb基因。在微氧条件下最大程度地诱导了启动子。通过PCR，SDS-PAGE和CO-差谱分析证实了表达的vgb的生化活性，所述CO-差谱分析显示了在420nm附近的特征吸收最大值。结果表明，vgb的表达可以促进细胞生长。研究了发酵培养基的组成和培养的物理条件。最佳培养基包括（g / L）：蔗糖30，豆粕5，NaNC_3 1，KH_2PO_4 1，K_2HPO_4 1.5，MgSO_4 0.6。确定了最佳发酵条件：pH 6.0，种子龄16 h，接种量10％，200 rpm，30°C，培养72 h，21后，vgb〜+重组体的最大结冷胶产量为1.713％+-0.02％。比亲本菌株高％。 VHb表现出对细胞生长的积极影响，并优化结冷胶产量。该结果提供了vgb基因在其他工业上重要菌株中的潜在应用，以解决多糖生产中需氧过程的不断增长的需求。

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