Eliminating host-guest incompatibility via enzyme mining enables the high-temperature production of N-acetylglucosamine

Wu, Yutong; Liu, Jiongqin; Han, Xiao; Meng, Xuanlin; Li, Mengke; Wang, Jing; Xue, Hongsong; Yang, Yuhan; Xu, Ping; Tao, Fei

doi:10.1016/j.isci.2022.105774

Cited by 7 publications

(4 citation statements)

References 74 publications

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“…To establish high-performance biocatalysis for Neu5Ac at high temperatures, the first step includes mining thermophilic AGEs and NanAs. Thermophilic AGEs were mined with our previously developed tool CEM () . Using the amino acid sequence of the well-characterized Anabaena sp.…”

Section: Resultsmentioning

confidence: 99%

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High-Performance Production of N-Acetyl-d-Neuraminic Acid with Whole Cells of Fast-Growing Vibrio natriegens via a Thermal Strategy

Peng,

Ma,

et al. 2023

J. Agric. Food Chem.

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Section: Resultsmentioning

confidence: 99%

“…Thermophilic AGEs were mined with our previously developed tool CEM (http://cem.sjtu.edu.cn). 42 Using the amino acid sequence of the well-characterized Anabaena sp. CH1 AGE (acAGE, GeneBank: ABG57043.1) as a query; Thermophagus xiamenensis AGE (txAGE, GeneBank: WP_010526912.1) was ultimately selected as the best candidate.…”

Section: ■ Results and Discussionmentioning

confidence: 99%

High-Performance Production of N-Acetyl-d-Neuraminic Acid with Whole Cells of Fast-Growing Vibrio natriegens via a Thermal Strategy

Peng,

Ma,

et al. 2023

J. Agric. Food Chem.

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“…The potential for high-temperature biomanufacturing has recently gained attention due to the discovery and application of several high-efficiency, thermophilic bacterial strains 15 , 40 , 41 . High temperatures similar to those used in chemical refineries enable faster fermentation, and thus shorter production cycles with reduced cooling costs and risk of contamination.…”

Section: Discussionmentioning

confidence: 99%

A thermostable type I-B CRISPR-Cas system for orthogonal and multiplexed genetic engineering

Yang,

Li,

et al. 2023

Nat Commun

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Thermophilic cell factories have remarkably broad potential for industrial applications, but are limited by a lack of genetic manipulation tools and recalcitrance to transformation. Here, we identify a thermophilic type I-B CRISPR-Cas system from Parageobacillus thermoglucosidasius and find it displays highly efficient transcriptional repression or DNA cleavage activity that can be switched by adjusting crRNA length to less than or greater than 26 bp, respectively, without ablating Cas3 nuclease. We then develop an orthogonal tool for genome editing and transcriptional repression using this type I-B system in both thermophile and mesophile hosts. Empowered by this tool, we design a strategy to screen the genome-scale targets involved in transformation efficiency and established dynamically controlled supercompetent P. thermoglucosidasius cells with high efficiency ( ~ 108 CFU/μg DNA) by temporal multiplexed repression. We also demonstrate the construction of thermophilic riboflavin cell factory with hitherto highest titers in high temperature fermentation by genome-scale identification and combinatorial manipulation of multiple targets. This work enables diverse high-efficiency genetic manipulation in P. thermoglucosidasius and facilitates the engineering of thermophilic cell factories.

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“…Therefore, using G. thermoglucosidasius as a chassis for fermentation production at high temperatures has practice prospects, which has the following advantages. This can reduce the cooling cost of large-scale exothermic fermentation, which becomes increasingly obvious with the increase in fermentation scale [24][25][26] . This reduces the risk of contamination by mesophilic microorganisms that cannot survive at high temperatures 27 .…”

Section: Introductionmentioning

confidence: 99%

Reprogramming a high robustGeobacillus thermoglucosidasiusfor efficient synthesis of polymer-grade lactic acid under extremely high temperature (60°C)

Liu

Han

Tao

et al. 2023

Preprint

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Optically pure lactic acid is an important precursor for the synthesis of biodegradable plastic polylactic acid. Geobacillus thermoglucosidasius is a thermophilic bacterial chassis with industrial potential. In this study, we reprogrammed this chassis to efficiently produce polymer-grade lactic acid by combining rational and semi-rational design strategies. We obtained optically pure L-lactic acid- and D-lactic acid-producing strains, GTD17 and GTD7, respectively, via rational strategies: constructing a lactic acid synthesis module, deleting by-product synthesis, and enhancing lactic acid production. At an extremely high temperature (60℃), the engineered strain GTD17 produced 94.2 g L−1 of L-lactic acid with an overall yield and productivity of 91.5% and 2.0 g L−1 h−1, respectively, and the optical purity of L-lactic acid was 99.5%. We then performed semi-rational adaptive evolution on the engineered strain GTD7 to obtain an optically pure D-lactic acid-producing strain GTD7-144; at the extremely high temperature (60℃), it produced 153.1 g L−1 of D-lactic acid with an overall yield and productivity of 93.0% and 3.2 g L−1 h−1, respectively. The optical purity of the D-lactic acid was 99.6%. Genome resequencing and analysis of the strain GTD7-144 revealed potential genetic targets that may improve production performance. This study represents an advancement in the high-temperature production of polymer-grade lactic acid by G. thermoglucosidasius.

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Eliminating host-guest incompatibility via enzyme mining enables the high-temperature production of N-acetylglucosamine

Cited by 7 publications

References 74 publications

High-Performance Production of N-Acetyl-d-Neuraminic Acid with Whole Cells of Fast-Growing Vibrio natriegens via a Thermal Strategy

High-Performance Production of N-Acetyl-d-Neuraminic Acid with Whole Cells of Fast-Growing Vibrio natriegens via a Thermal Strategy

A thermostable type I-B CRISPR-Cas system for orthogonal and multiplexed genetic engineering

Reprogramming a high robustGeobacillus thermoglucosidasiusfor efficient synthesis of polymer-grade lactic acid under extremely high temperature (60°C)

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