Effect of different sucrose concentrations on the biosynthesis of poly-3-hydroxybutyrate and alginate by the bacterial strain Azotobacter vinelandii 12 under different aeration conditions

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Abstract

Bacteria of the genus Azotobacter sp. produce two classes of biologically important biocompatible and biodegradable polymers – polyoxyalkanoates, which are the bacterial reserve, and alginates (ALG), which perform the function of protecting nitrogenase from oxygen. Both polymers are becoming increasingly important for use in bioengineering, pharmaceuticals and medicine, so studies of their biosynthesis and properties are currently highly relevant. The present work shows the possibility of regulating alginate and poly-3-hydroxybutyrate (PHB) synthesis by A. vinelandii 12 culture depending on the increase of sucrose concentration in the medium under different aeration conditions. At high aeration and high sucrose concentration in the medium (50 g/L), the maximum yield of free (1.08 g/L) and capsular ALG (2.26 g/L) in the medium was obtained. Under low aeration conditions, the synthesis of free ALG was completely inhibited. The maximum value of РНB synthesis was observed at medium aeration and high concentration of sucrose (50 g/l) in the medium. The maximum molecular weight (MW) of ALG was 477 kDa, while the maximum MW of PHB was much higher, reaching 1479 kDa. At low sucrose concentrations in the medium (5 to 20 g/l), capsular ALG is predominantly synthesized (up to 100% of the sum of all polymers) at all aeration levels. With increasing sucrose concentration, PHB is predominantly synthesized (68%) under low aeration conditions, an equal ratio of PHB and capsular ALG synthesis is observed under medium aeration conditions, and free ALG is actively synthesized under high aeration conditions. This work demonstrates the possibility of obtaining a selective synthesis of ALG or PHB by A. vinelandii 12 by modifying its cultivation conditions. The results obtained can be used for the development of directed biosynthesis of target products (PHB and ALG) in biotechnology.

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About the authors

A. A. Dudun

Research Center of Biotechnology of the Russian Academy of Sciences; Gamaleya National Research Centre for Epidemiology and Microbiology, Ministry of Health of the Russian Federation

Author for correspondence.
Email: dudunandrey@mail.ru

Bach Institute of Biochemistry

Russian Federation, Moscow, 119071; Moscow, 123098

T. K. Makhina

Research Center of Biotechnology of the Russian Academy of Sciences

Email: dudunandrey@mail.ru

Bach Institute of Biochemistry

Russian Federation, Moscow, 119071

A. P. Bonartsev

M.V. Lomonosov Moscow State University

Email: dudunandrey@mail.ru

Faculty of Biology

Russian Federation, Moscow, 119234

G. A. Bonartseva

Research Center of Biotechnology of the Russian Academy of Sciences

Email: dudunandrey@mail.ru

Bach Institute of Biochemistry

Russian Federation, Moscow, 119071

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Supplementary files

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2. Fig. 1. Growth of the bacterial strain A. vinelandii 12 at different levels of aeration and sucrose concentration in the medium from 5 g/l to 50 g/l (fermentation time – 72 h): 1 – 150 rpm; 2 – 200 rpm; 3 – 250 rpm. Multiple comparison according to Kruskal-Wallis between all groups has a significance level of p < 0.05.

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3. Fig. 2. Biosynthesis of free (a, g/l) and capsular (b, g/l) ALG, productivity of Azotobacter vinelandii 12 synthesis of free (c) and capsular (d) ALG: 1 – 150 rpm; 2 – 200 rpm; 3 – 250 rpm. Multiple comparison according to Kruskal-Wallis between all groups has a significance level of p < 0.05.

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4. Fig. 3. Biosynthesis (a, g/l) and productivity (b, g/g) of PHB by the bacterial strain A. vinelandii 12: 1 – 150 rpm; 2 – 200 rpm; 3 – 250 rpm. Multiple comparison according to Kruskal-Wallis between all groups has a significance level of p < 0.05.

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5. Fig. 4. MM of capsular ALGs synthesized at high aeration levels by Azotobacter vinelandii 12 bacteria and different sucrose concentrations. Kruskal-Wallis multiple comparison between all groups has a significance level of p < 0.05.

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6. Fig. 5. Composition of synthesized biopolymers (PHB and ALG) by Azotobacter vinelandii 12 cells at different sucrose concentrations in the medium and different aeration: a – 150 rpm; b – 200 rpm; c – 250 rpm.

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