Departamento de Ingeniería Celular y Biocatálisis, Instituto de Biotecnología, Universidad Nacional Autónoma de México, Av. Universidad 2001, Col. Chamilpa, Cuernavaca, 62210, Morelos, Mexico.
Enzyme Microb Technol. 2013 Jul 10;53(2):85-91. doi: 10.1016/j.enzmictec.2013.04.010. Epub 2013 May 14.
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.
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。