Optimization of medium compositions to improve a novel glycoprotein production by Streptomyces kanasenisi ZX01

Abstract

Streptomyces kanasenisi ZX01 was found to produce a novel glycoprotein GP-1 previously, which was secreted into medium and had significant activity against tobacco mosaic virus. However, the low production of GP-1 by strain ZX01 limited its further studies. In order to improve the yield of GP-1, a series of statistical experimental design methods were applied to optimize medium of strain ZX01 in this work. Millet medium was chosen to be the optimal original medium for optimization. Soluble starch and yeast extract were identified as the optimal carbon and nitrogen source using one-factor-at-a-time method. Response surface methodology was used to optimize medium compositions (soluble starch, yeast extract and inorganic salts). A higher yield of GP-1 was 601.33 µg/L after optimization. The optimal compositions of medium were: soluble starch 13.61 g/L, yeast extract 4.19 g/L, NaCl 3.54 g/L, CaCO3 0.28 g/L, millet, 10 g/L. The yield of GP-1 in a 5 L fermentor using optimized medium was 2.54 mg/L, which is much higher than the result of shake flask. This work will be helpful for the improvement of GP-1 production on a large scale and lay a foundation for developing it to be a novel anti-plant virus agent.

Effects of Agitation, Aeration and Temperature on Production of a Novel Glycoprotein GP-1 by Streptomyces kanasenisi ZX01 and Scale-Up Based on Volumetric Oxygen Transfer Coefficient

“…The results reveal that under the optimal conditions, the final GP-1 production and the maximum DCW could reach 3.92 mg/L and 3.31 g/L on average. The final GP-1 production of 3.92 mg/L on a bench scale was increased by 54.33% compared with 2.54 mg/L before optimization [ 35 ]. On the other side, average value of k L a was 21.62 h −1 , which was an important parameter for scale-up fermentation from bench scale to pilot scale.…”

“…Our previous study indicated that strain ZX01 can produce a novel glycoprotein GP-1 with activity against plant viruses, especially tobacco mosaic virus (TMV) [ 33 , 34 ]. However, batch fermentation of strain ZX01 was found to be extremely unstable, affording low GP-1 production and requiring long fermentation periods, which not only reduce tremendously the efficiency of future research, but also seriously hinder potential industrial production of the strain and commercial development of its related fermentation products [ 35 ].…”

Optimization of Production Conditions for Protoplasts and Polyethylene Glycol-Mediated Transformation of Gaeumannomyces tritici

“…The fit of the model was checked using the determination coefficient, R 2 , which was 0.9812, indicating that 98% of the variability in the response could be explained by the model. The coefficient of variation (CV) was related to the reliability and precision, and the higher the value of CV, the lower the reliability of the experiment [ 24 ]. In this model, a lower CV value (3.89%) indicated better reliability and precision of the experiment.…”

Optimization of Fermentation Conditions and Bench-Scale for Improvement of a Novel Glycoprotein GP-1 Production by Streptomyces kanasenisi ZX01

“…This has tremendous potential to become a novel anti-TMV agent with low toxicity, a broad spectrum, and high environmental compatibility [ 13 ]. However, extremely low fermentation production has largely hindered further research and market applications of strain ZX01, glycoprotein GP-1 and other metabolites [ 14 ]. For the above reason, the purpose of this work was to improve glycoprotein GP-1 production through optimizing the fermentation conditions (inoculum volume, initial pH and rotating speed) using response surface methodology.…”

“…Glycoprotein GP-1, with the molecular weight of approximately 8.5 kDa, contains 40.23% carbohydrates and 54.36% polypeptides with N-linked and O-linked glycan [ 12 , 13 ]. Our previous study indicated that glycoprotein GP-1 production was strongly affected by medium components, especially the types and concentrations of carbon and nitrogen sources [ 14 ]. Process factors (temperature, pH, rotating speed) also play an important role in improving objective metabolites production and may contribute to the reduction of fermentation cost, even changing the path of microbial biosynthesis to discover new natural products [ 15 , 16 , 17 ].…”