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26. A cell-free artificial anabolic pathway for direct conversion of CO2 to ethanol.

Author

Dong, Wanrong, Ji, Xiuling, Huang, Yuhong, Xue, Yaju, Guo, Boxia, Cai, Dongbo, Chen, Shouwen, and Zhang, Suojiang

Subjects
ETHANOL, ENZYME activation, CARBON cycle, ACETYLCOENZYME A, MODULAR design, FORMALDEHYDE
Abstract

Biological CO2 activation and conversion to high-value ethanol are a feasible and green strategy to close the carbon cycle. However, naturally evolved CO2 utilization pathways involve carbon loss or ATP consumption. Herein, we report a complete anabolic pathway for direct conversion of CO2 to ethanol by constructing and assembling three functional modules including CO2 activation, formaldehyde → acetyl-CoA, and ethanol synthesis in a carbon-conserved and ATP-independent system. These artificial ethanol metabolic pathway fluxes were strengthened by screening efficient key enzymes for CO2 activation, promoting formaldehyde assimilation, and employing a two-step reaction. 13C-labeled CO2 demonstrated the feasibility of the pathway converting CO2 into ethanol in vitro by detecting the carbon flow. With this anabolic pathway, we obtained an ethanol concentration of 0.37 mM at a conversion rate of 4.33 nmol CO2 min−1 mg−1 enzyme and 0.5 mM R5P supply. This modularization strategy provides a new avenue in the construction of artificial metabolic pathways for ethanol synthesis from CO2. [ABSTRACT FROM AUTHOR]

Published
2023
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27. Systematic metabolic engineering of Bacillus licheniformis for hyperproduction of the antioxidant hydroxytyrosol.

Author

Zhan, Yangyang, Zhou, Fei, Ruan, Wenqi, Yin, Hao, Li, Zhi, Wang, Huan, Li, Tao, Cai, Dongbo, Yang, Shihui, Ma, Xin, and Chen, Shouwen

Subjects
BACILLUS licheniformis, HYDROXYTYROSOL, ALCOHOL dehydrogenase, ANTIOXIDANTS, PRODUCTION increases
Abstract

Hydroxytyrosol (HT) is a natural antioxidant that exhibits various health-related properties. In this work, we focused on establishing and optimizing the production of HT in Bacillus licheniformis. Through protein engineering ketoacid decarboxylase, boosting the phosphoenolpyruvate (PEP) supply, releasing feedback inhibition, and blocking competing pathways, HT production increased to 2078 mg L−1 from zero. Remarkably, the tyrosol catabolic pathway was identified and deletion of the alcohol dehydrogenase gene adhA significantly reduced tyrosol degradation. Next, multiple rate-liming factors were identified, including insufficiency of PEP, erythrose-4-phosphate and 4-hydroxyphenylpyruvate supply, and imbalance of KivDV461I and HpaBC expression levels. The carbon flux toward HT was significantly improved by modulating the glucose transport system, fine-tuning the expression levels of kivDV461I and hpaBC, and enhancing the expression of tyrAfbr, tkt, and zwf, which increased the HT titer to 6299 mg L−1. Finally, the final strain DHT23 produced 9475 mg L−1 HT under fed-batch conditions, with a yield and productivity of 135.4 mg g−1 glucose and 395 mg (L h)−1, respectively, which represents the highest HT production to date. Taken together, this study establishes B. licheniformis as a framework for HT hyperproduction and paves the way for achieving the industrial production of HT in the future. [ABSTRACT FROM AUTHOR]

Published
2023
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28. Point Cloud-Based Smart Building Acceptance System for Surface Quality Evaluation.

Author

Cai, Dongbo, Chai, Shaoqiang, Wei, Mingzhuan, Wu, Hui, Shen, Nan, Zhou, Yin, Ding, Yanchao, Hu, Kaixin, and Hu, Xingyi

Subjects
BUILDING additions, POINT cloud, INTELLIGENT buildings, MANUAL labor, ACCURACY of information, INFORMATION storage & retrieval systems
Abstract

The current expansion of building structures has created a demand for efficient and smart surface quality evaluation at the acceptance phase. However, the conventional approach mainly relies on manual work, which is labor-intensive, time-consuming, and unrepeatable. This study presents a systematic and practical solution for surface quality evaluation of indoor building elements during the acceptance phase using point cloud. The practical indoor scanning parameters determination procedure was proposed by analyzing the project requirements, room environment, and apparatus. An improved DBSCAN algorithm was developed by introducing a plane validation and coplanar checking to facilitate the surface segmentation from the point cloud. And a revised Least Median of Square-based algorithm was proposed to identify the best-fit plane. Afterwards, the flatness, verticality, and squareness were evaluated and depicted using a color-coded map based on the segmented point cloud. The experiment on an apartment showcases how the system improves the information flow and accuracy during building acceptance, resulting in a potentially smart acceptance activity. [ABSTRACT FROM AUTHOR]

Published
2023
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41. Additional file 1 of Enhancement of alkaline protease production in recombinant Bacillus licheniformis by response surface methodology

Author

Zhang, Ying, Hu, Jingmin, Zhang, Qing, Cai, Dongbo, Chen, Shouwen, and Wang, Yonghong

Abstract

Additional file 1: Table S1. Factors and levels of Plackett-Burman experiment. Table S2. Factors and levels of Central Composite design. Table S3. Plackett-Burma test design and results. Table S4. The variance analysis of Plackett-Burman test. Table S5. The design and results of steepest ascent experiment. Table S6. CCD design and results. Table S7. The variance analysis of experiment. Figure S1. The alkaline protease and process pH during the fermentation of different initial medium pH.

Published
2023
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50. Additional file 1 of Minimization and optimization of α-amylase terminator for heterologous protein production in Bacillus licheniformis

Author

Rao, Yi, Yang, Jingyao, Wang, Jiaqi, Yang, Xinyuan, Zhang, Mengxi, Zhan, Yangyang, Ma, Xin, Cai, Dongbo, Wang, Zhangqian, and Chen, Shouwen

Abstract

Additional file1: Table S1. Strains and plasmids used in this study. Table S2. Primers used in this study. Table S3. The effects of GC content at the bottom of stem-loop structure, Gibbs free energy ΔG, U-tract length on terminator performances. Table S4. The sequences of different terminators plasmids used in this study. Table S5. Comparison of termination efficiency of different terminators in Bacillus. Figure S1. The secondary structure map of TamyL with a size of 501 bp. Figure S2. The construction map of plasmids with different terminators and dual reporter genes.

Published
2022
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