Engineering the methyltransferase through inactivation of the genK and genL leads to a significant increase of gentamicin C1a production in an industrial strain of Micromonospora echinospora 49-92S

Xu, Feng; Zhang, Xinyu; Liu, Ling; Ke, Xiang; Wu, Jie; Guo, Yuanxin; Tian, Xiaohui; Chu, Ju

doi:10.1007/s00449-022-02774-0

Cited by 4 publications

(4 citation statements)

References 26 publications

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“…In addition, the concentration of NH 4 + was determined using a Roche biochemical analyzer (Roche Inc.), while Ca 2+ was measured using a Siemens serum analyzer (SIEMENS AG). The DNS method that commonly used in the fermentation process was employed to determine the total sugar concentration (Xu et al, 2022), which includes soluble starch. During the fermentation process, it could be found that the consumption rate of total sugar in the fermentation broth varies under different pH conditions, indicating that C. Butyricum has the ability to produce amylase, and therefore can utilize starch as a carbon source for cell growth and spore production.…”

Section: Methodsmentioning

confidence: 99%

Enhancing and monitoring spore production in Clostridium butyricum using pH‐based regulation strategy and a robust soft sensor based on back‐propagation neural networks

Xu,

Zhang,

Wang

et al. 2023

Biotech & Bioengineering

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Clostridium butyricum (C. butyricum) is a probiotic that forms anaerobic spores and plays a crucial role in regulating gut microbiota. However, the total viable cell count and spore yield of C. butyricum in industrial production are comparatively low. To this end, we investigated the metabolic characteristics of the strain and proposed three distinct pH regulation strategies for enhancing spore production. In addition, precise measurement of fermentation parameters such as substrate concentration, total viable cell count, and spore concentration is crucial for successful industrial probiotics production. Nevertheless, online measurement of these intricate parameters in the fermentation of C. butyricum poses a considerable challenge owing to the complex, nonlinear, multivariate, and strongly coupled characteristics of the production process. Therefore, we analyzed the capacitance and conductivity acquired from a viable cell sensor as the core parameters for the fermentation process. Subsequently, a robust soft sensor was developed using a seven‐input back‐propagation neural network model with input variables of fermentation time, capacitance, conductivity, pH, initial total sugar concentration, ammonium ion concentration, and calcium ion concentration. The model enables the online monitoring of total viable biomass count, substrate concentrations, and spore yield, and can be extended to similar fermentation processes with pH changes as a characteristic feature.This article is protected by copyright. All rights reserved.

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Section: Methodsmentioning

confidence: 99%

Enhancing and monitoring spore production in Clostridium butyricum using pH‐based regulation strategy and a robust soft sensor based on back‐propagation neural networks

Xu,

Zhang,

Wang

et al. 2023

Biotech & Bioengineering

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“…The industrial production of etimicin requires a significant quantity of high-purity gentamicin C1a. As a result, many researchers have employed genetic engineering techniques to develop strains that produce high yields of single-component C1a [6,[19][20][21].…”

Section: Determination Of Sugar Amino Nitrogen Germ Concentration And...mentioning

confidence: 99%

“…Gentamicin biosynthesis is not only related to the biosynthesis and assembly of gentamicin but also to the metabolism of carbohydrates [6]. This is mainly because glucose-6-phosphate is an important precursor of gentamicin (Figure 1).…”

Section: Gentamicin Biosynthetic Pathway Analysismentioning

confidence: 99%

“…It contains a 2-deoxystreptamine (2-DOS) ring on C4 and C6, which is connected through glycosidic bonds to garosamine and purpurosamine. The majority of the enzymes used in the synthesis of gentamicin have been extensively elucidated [5][6][7][8][9][10][11][12][13]. The process of synthesizing gentamicin begins with D-glucose-phosphate, which is converted into 2-DOS by the isomerase GenC, the aminotransferase GenS1, and the dehydrogenase GenE.…”

Section: Introductionmentioning

confidence: 99%

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Proteomic Analysis of the Effect of CaCl2 and Sodium Citrate on Gentamicin Biosynthesis of Micromonospora echinospora SIPI-GM.01

Yang,

Lin,

et al. 2023

Fermentation

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The clinical antibiotic gentamicin is a mixture of several difficult-to-separate components, the minor group of which is gentamicin C1a, a precursor for the synthesis of the high-efficacy and low-toxicity antibiotic etimicin. This study aimed to achieve the high production of gentamicin as well as gentamicin C1a. In this study, the influence of organic and inorganic salts on the gentamicin production was screened and label-free proteomics was used to determine the mechanisms responsible for the effects. In 25 L fermentation experiments, the addition of 0.1% CaCl2 and 0.3% sodium citrate increased gentamicin titers by 11.5% (2398 μg/mL vs. 2150 μg/mL), while the C1a ratio increased from 38% to 42%. The results showed that CaCl2 downregulated the synthesis and metabolism of the tetrapyrrole pathway and the GenK protein (0.08-fold) in the gentamicin synthesis pathway, whereas sodium citrate downregulated key proteins in the glycosylation pathway and tricarboxylic acid pathway. Thus, CaCl2 caused changes in methylation during the synthesis of gentamicin, increasing the proportion of gentamicin C1a. In contrast, sodium citrate inhibited primary metabolism to promote the production of secondary metabolites of gentamicin. This study provided a basis for the co-production of gentamicin C1a mono-component and gentamicin multicomponent.

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Ammonium sulfate supplementation enhances erythromycin biosynthesis by augmenting intracellular metabolism and precursor supply in Saccharopolyspora erythraea

Yuan

et al. 2023

Bioprocess Biosyst Eng

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Engineering the methyltransferase through inactivation of the genK and genL leads to a significant increase of gentamicin C1a production in an industrial strain of Micromonospora echinospora 49-92S

Cited by 4 publications

References 26 publications

Enhancing and monitoring spore production in Clostridium butyricum using pH‐based regulation strategy and a robust soft sensor based on back‐propagation neural networks

Enhancing and monitoring spore production in Clostridium butyricum using pH‐based regulation strategy and a robust soft sensor based on back‐propagation neural networks

Proteomic Analysis of the Effect of CaCl2 and Sodium Citrate on Gentamicin Biosynthesis of Micromonospora echinospora SIPI-GM.01

Ammonium sulfate supplementation enhances erythromycin biosynthesis by augmenting intracellular metabolism and precursor supply in Saccharopolyspora erythraea

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