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在粘细菌中,algD 基因和 algE 基因分别编码 alginate lyase 和 alginate polymerase,algR 基因编码调控蛋白 AlgR。AlgR 蛋白与 algD 基因上游的 algR 操纵子结合,正向调控 algD 基因的转录。algE 基因的转录则受到负调控,algR 蛋白与 algE 基因上游的 algE 操纵子结合,抑制 algE 基因的转录。algD 基因和 algE 基因的转录均受到 AlgR 蛋白的调控。

Expression of alginases and alginate polymerase genes in response to oxygen, and their relationship with the alginate molecular weight in Azotobacter vinelandii.

机构信息

Departamento de Ingeniería Celular y Biocatálisis, Instituto de Biotecnología, Universidad Nacional Autónoma de México, Av. Universidad 2001, Col. Chamilpa, Cuernavaca, 62210, Morelos, Mexico.

出版信息

Enzyme Microb Technol. 2013 Jul 10;53(2):85-91. doi: 10.1016/j.enzmictec.2013.04.010. Epub 2013 May 14.

Abstract

The transcription of genes involved in alginate polymerization and depolymerization, as well as the alginase activity (extracellular and intracellular) under oxygen-limited and non oxygen-limited conditions in cultures of A. vinelandii, was studied. Two levels of dissolved oxygen tension (DOT) (1% and 5%, oxygen-limited and non-oxygen-limited, respectively) strictly controlled by gas blending, were evaluated in a wild type strain. In cultures at low DOT (1%), in which a high molecular weight alginate (1200 kDa) was synthesized, the transcription levels of alg8 and alg44 (genes encoding alginate polymerase complex), and algX (encoding a protein involved in polymer transport through periplasmic space) were considerably higher as compared to cultures conducted at 5% DOT, under which an alginate with a low MW (42 kDa) was produced. In the case of genes encoding for intracellular and extracellular alginases, the levels of these transcripts were higher at 1% DOT. However, intracellular and extracellular alginase activity were lower (0.017 and 0.01 U/mg protein, respectively) in cultures at 1% DOT, as compared with the activities measured at 5% DOT (0.027 and 0.052 U/mg protein for intracellular and extracellular maximum activity, respectively). The low alginase activity measured in cultures at 1% DOT and the high level of transcription of genes constituting alginate polymerase complex might be mechanisms by which oxygen regulates the production of alginates with a high MW.

摘要

研究了 Alginate 聚合和降解相关基因的转录,以及在缺氧和非缺氧条件下,A. vinelandii 培养物中的 Alginate 酶活性(胞外和胞内)。通过气体混合严格控制了两种溶解氧张力(DOT)水平(1%和 5%,分别为缺氧和非缺氧),并在野生型菌株中进行了评估。在低 DOT(1%)的培养物中,合成了高分子量 Alginate(1200 kDa),与在 5% DOT 下培养的 Alginate 相比,Alg8 和 Alg44(编码 Alginate 聚合酶复合物的基因)以及 AlgX(编码一种参与聚通过周质空间运输的蛋白质)的转录水平要高得多。对于编码胞内和胞外 Alginate 酶的基因,在 1% DOT 下,这些转录本的水平更高。然而,在 1% DOT 的培养物中,胞内和胞外 Alginate 酶活性较低(分别为 0.017 和 0.01 U/mg 蛋白),而在 5% DOT 下测量的活性较低(分别为 0.027 和 0.052 U/mg 蛋白,为胞内和胞外最大活性)。在 1% DOT 的培养物中测量的 Alginate 酶活性较低,以及构成 Alginate 聚合酶复合物的基因的高水平转录,可能是氧气调节高分子量 Alginate 产生的机制。

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