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通过基因修饰和嗜酸乳杆菌培养基优化提高透明质酸产量。

Hyaluronic acid production enhancement via genetically modification and culture medium optimization in Lactobacillus acidophilus.

机构信息

National Institute for Genetic Engineering and Biotechnology (NIGEB), Institute of Industrial and Environmental Biotechnology, Bioprocess Engineering Research Group, Shahrak-e Pajoohesh km 15, Tehran-Karaj Highway, P. O. Box: 14965/161, Tehran, Iran; Department of Biology, Faculty of Sciences, University of Zanjan, Iran.

National Institute for Genetic Engineering and Biotechnology (NIGEB), Institute of Industrial and Environmental Biotechnology, Bioprocess Engineering Research Group, Shahrak-e Pajoohesh km 15, Tehran-Karaj Highway, P. O. Box: 14965/161, Tehran, Iran.

出版信息

Int J Biol Macromol. 2019 Jan;121:870-881. doi: 10.1016/j.ijbiomac.2018.10.112. Epub 2018 Oct 17.

Abstract

Hyaluronic acid (HA) is a natural polymer with various molecular weights that specify multiple biological roles. Traditionally, HA is obtained from animal waste and conventional pathogenic streptococci. However, there are challenges in these processes such as the presence of exotoxins, hyaluronidase, and viral contamination. In order to reduce these problems, this study was conducted to produce HA using recombinant bacterium that is generally recognized as safe (GRAS), and thereafter increase production through experimental design. At first, some lactic acid bacteria were screened and evaluated for HA production. Accordingly, among the selected bacteria, Lactobacillus acidophilus PTCC1643 produced about 0.25 g HA/L in the 48th hour of cultivation, and was thus selected as an alternative host for heterologous HA production. An expression vector containing HA synthase genes was transformed into L. acidophilus by electroporation. Consequently, HA production increased to 0.4 g/L. Eventually, response surface method (RSM) was used, which increased HA production to 1.7 g/L. This is approximately 7-fold higher than that produced at first. The resulting HA was characterized by FTIR spectroscopy and its molecular weight was estimated using agarose gel electrophoresis. In conclusion, L. acidophilus could be a safe, effective, and novel HA producer with industrial potential and commercial prospects.

摘要

透明质酸(HA)是一种具有多种分子量的天然聚合物,具有多种生物学功能。传统上,HA 是从动物废物和常规致病性链球菌中获得的。然而,这些过程存在一些问题,如存在外毒素、透明质酸酶和病毒污染。为了减少这些问题,本研究使用普遍认为安全(GRAS)的重组细菌生产 HA,并通过实验设计提高产量。首先,筛选并评估了一些乳酸菌生产 HA 的能力。因此,在选择的细菌中,嗜酸乳杆菌 PTCC1643 在培养的第 48 小时产生约 0.25g/L 的 HA,因此被选为异源 HA 生产的替代宿主。通过电穿孔将含有 HA 合酶基因的表达载体转化为嗜酸乳杆菌。结果,HA 的产量增加到 0.4g/L。最终,采用响应面法(RSM),HA 的产量增加到 1.7g/L。这大约是最初产量的 7 倍。用傅里叶变换红外光谱法(FTIR)对所得 HA 进行了表征,并通过琼脂糖凝胶电泳法估计了其分子量。总之,嗜酸乳杆菌可能是一种具有工业潜力和商业前景的安全、有效、新型的 HA 生产菌。

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