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学者姓名：范立海
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Abstract ：
(D)-Glucaric acid is a value-added dicarboxylic acid that can be utilized in the chemical, food, and pharmaceutical industries. Due to the complex process and environmental pollution associated with the chemical production of d-glucaric acid, bioconversion for its synthesis has garnered increasing attention in recent years. In this study, a novel cell factory was developed for the efficient production of d-glucaric acid using d-xylose and methanol. Mdh, Hps, Phi, Miox, Ino1, Suhb, and Udh were first co-expressed in E. coli JM109 to construct the d-glucaric acid synthesis pathway. The deletion of FrmRAB, RpiA, PfkA, and PfkB was then performed to block or weaken the endogenous competitive pathways. Next, adaptive evolution was carried out to improve cell growth and substrate utilization. With the purpose of further increasing the product titer, the NusA tag and myo-inositol biosensor were introduced into engineered E. coli to enhance Miox expression. After medium optimization and fermentation process control, 3.0 g/L of d-glucaric acid was finally obtained in the fed-batch fermentation using modified Terrific Broth medium.
Keyword ：
d-glucaric acid d-glucaric acid Escherichia coli Escherichia coli metabolic engineering metabolic engineering microbial fermentation microbial fermentation
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| GB/T 7714 | Chen, Wei-Xiang , Zheng, Ling-Jie , Luo, Xuan et al. Metabolic Engineering and Adaptive Evolution of Escherichia coli for Enhanced Conversion of D-Xylose to D-Glucaric Acid Mediated by Methanol [J]. | BIOTECHNOLOGY AND BIOENGINEERING , 2025 , 122 (6) : 1472-1483 . |
| MLA | Chen, Wei-Xiang et al. "Metabolic Engineering and Adaptive Evolution of Escherichia coli for Enhanced Conversion of D-Xylose to D-Glucaric Acid Mediated by Methanol" . | BIOTECHNOLOGY AND BIOENGINEERING 122 . 6 (2025) : 1472-1483 . |
| APA | Chen, Wei-Xiang , Zheng, Ling-Jie , Luo, Xuan , Zheng, Shang-He , Zheng, Hui-Dong , Fan, Li-Hai et al. Metabolic Engineering and Adaptive Evolution of Escherichia coli for Enhanced Conversion of D-Xylose to D-Glucaric Acid Mediated by Methanol . | BIOTECHNOLOGY AND BIOENGINEERING , 2025 , 122 (6) , 1472-1483 . |
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Abstract ：
d-Allulose, a C-3 epimer of d-fructose, has great market potential in food, healthcare, and medicine due to its excellent biochemical and physiological properties. Microbial fermentation for d-allulose production is being developed, which contributes to cost savings and environmental protection. A novel metabolic pathway for the biosynthesis of d-allulose from a d-xylose-methanol mixture has shown potential for industrial application. In this study, an artificial antisense RNA (asRNA) was introduced into engineered Escherichia coli to diminish the flow of pentose phosphate (PP) pathway, while the UDP-glucose-4-epimerase (GalE) was knocked out to prevent the synthesis of byproducts. As a result, the d-allulose yield on d-xylose was increased by 35.1%. Then, we designed a d-xylose-sensitive translation control system to regulate the expression of the formaldehyde detoxification operon (FrmRAB), achieving self-inductive detoxification by cells. Finally, fed-batch fermentation was carried out to improve the productivity of the cell factory. The d-allulose titer reached 98.6 mM, with a yield of 0.615 mM/mM on d-xylose and a productivity of 0.969 mM/h. © 2024 American Chemical Society.
Keyword ：
d-allulose d-allulose Escherichia coli Escherichia coli fermentation fermentation metabolic engineering metabolic engineering
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| GB/T 7714 | Guo, Q. , Zheng, L.-J. , Zheng, S.-H. et al. Enhanced Biosynthesis of d-Allulose from a d-Xylose-Methanol Mixture and Its Self-Inductive Detoxification by Using Antisense RNAs in Escherichia coli [J]. | Journal of Agricultural and Food Chemistry , 2024 , 72 (26) : 14821-14829 . |
| MLA | Guo, Q. et al. "Enhanced Biosynthesis of d-Allulose from a d-Xylose-Methanol Mixture and Its Self-Inductive Detoxification by Using Antisense RNAs in Escherichia coli" . | Journal of Agricultural and Food Chemistry 72 . 26 (2024) : 14821-14829 . |
| APA | Guo, Q. , Zheng, L.-J. , Zheng, S.-H. , Zheng, H.-D. , Lin, X.-C. , Fan, L.-H. . Enhanced Biosynthesis of d-Allulose from a d-Xylose-Methanol Mixture and Its Self-Inductive Detoxification by Using Antisense RNAs in Escherichia coli . | Journal of Agricultural and Food Chemistry , 2024 , 72 (26) , 14821-14829 . |
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Abstract ：
D-allulose, a naturally occurring monosaccharide, is present in small quantities in nature. It is considered a valuable low-calorie sweetener due to its low absorption in the digestive tract and zero energy for growth. Most of the recent efforts to produce D-allulose have focused on in vitro enzyme catalysis. However, microbial fermentation is emerging as a promising alternative that offers the advantage of combining enzyme manufacturing and product synthesis within a single bioreactor. Here, a novel approach was proposed for the efficient biosynthesis of D-allulose from glycerol using metabolically engineered Escherichia coli. FbaA, Fbp, AlsE, and A6PP were used to construct the D-allulose synthesis pathway. Subsequently, PfkA, PfkB, and Pgi were disrupted to block the entry of the intermediate fructose-6-phosphate (F6P) into the Embden−Meyerhof−Parnas (EMP) and pentose phosphate (PP) pathways. Additionally, GalE and FryA were inactivated to reduce D-allulose consumption by the cells. Finally, a fed-batch fermentation process was implemented to optimize the performance of the cell factory. As a result, the titer of D-allulose reached 7.02 g/L with a maximum yield of 0.287 g/g. © 2024 Elsevier B.V.
Keyword ：
D-allulose D-allulose Escherichia coli Escherichia coli Fermentation Fermentation Metabolic engineering Metabolic engineering
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| GB/T 7714 | Guo, Q. , Dong, Z.-X. , Luo, X. et al. Engineering Escherichia coli for D-allulose biosynthesis from glycerol [J]. | Journal of Biotechnology , 2024 , 394 : 103-111 . |
| MLA | Guo, Q. et al. "Engineering Escherichia coli for D-allulose biosynthesis from glycerol" . | Journal of Biotechnology 394 (2024) : 103-111 . |
| APA | Guo, Q. , Dong, Z.-X. , Luo, X. , Zheng, L.-J. , Fan, L.-H. , Zheng, H.-D. . Engineering Escherichia coli for D-allulose biosynthesis from glycerol . | Journal of Biotechnology , 2024 , 394 , 103-111 . |
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Abstract ：
制备Pd负载在氮掺杂碳纳米管(NCNTs)载体上的Pd/NCNTs催化剂，并将其用于间二硝基苯催化加氢生成间苯二胺的反应。研究NCNTs的不同制备方法对Pd/NCNTs催化剂催化性能的影响。结果表明，Pd纳米颗粒可以更均匀地分散在NCNTs载体上，可与载体形成强相互作用。当NCNTs中的氮含量越高且吡啶氮占比较高时，Pd/NCNTs催化剂的加氢活性越高。在间二硝基苯催化加氢反应中，Pd/NCNTs催化剂相较于Pd/CNTs催化剂的加氢活性提高了69%。
Keyword ：
催化剂工程 催化剂工程 吡啶氮 吡啶氮 氮掺杂 氮掺杂 硝基加氢 硝基加氢 钯催化剂 钯催化剂 间二硝基苯 间二硝基苯
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| GB/T 7714 | 刘加帅 , 王楠 , 郑辉东 et al. 氮掺杂碳纳米管负载钯基催化剂在间二硝基苯加氢中的应用 [J]. | 工业催化 , 2024 , 32 (08) : 24-30 . |
| MLA | 刘加帅 et al. "氮掺杂碳纳米管负载钯基催化剂在间二硝基苯加氢中的应用" . | 工业催化 32 . 08 (2024) : 24-30 . |
| APA | 刘加帅 , 王楠 , 郑辉东 , 范立海 , 吴丹 . 氮掺杂碳纳米管负载钯基催化剂在间二硝基苯加氢中的应用 . | 工业催化 , 2024 , 32 (08) , 24-30 . |
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Abstract ：
间氨基苯磺酸钠是一种重要的化工中间体,可由多种方法制备得到,催化加氢法是目前主要使用的方法,但工业化报道较少,这是由于在间硝基苯磺酸钠加氢合成间氨基苯磺酸钠的液相反应中催化剂存在失活现象,难以多次套用,工业化成本高.使用自制Pd/C催化剂进行间硝基苯磺酸钠的液相加氢反应,采用液相色谱-四级杆飞行时间串联质谱(LC-QTOF-MS)、X射线衍射谱(XRD)、X射线光电子能谱(XPS)、BET表面分析等方法对Pd/C催化剂的失活原因进行探究,确定催化剂的失活原因.Pd/C催化剂的失活是由于在硝基选择性加氢过程中,原料间硝基苯磺酸钠转化为偶氮化合物或氧化偶氮化合物,由于这些中间体的吸附性较强,易堵塞活性炭载体的孔道,造成催化剂比表面积减少,导致催化剂暂时性的中毒失活,而并非是加氢还原了原料中磺酸基团生成硫化钯而导致的催化剂永久性失活.
Keyword ：
Pd/C催化剂 Pd/C催化剂 催化剂失活 催化剂失活 催化加氢 催化加氢 催化化学 催化化学 间硝基苯磺酸钠 间硝基苯磺酸钠
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| GB/T 7714 | 林知滨 , 郑辉东 , 范立海 et al. Pd/C催化剂在间硝基苯磺酸钠加氢反应中的应用 [J]. | 工业催化 , 2024 , 32 (10) : 56-62 . |
| MLA | 林知滨 et al. "Pd/C催化剂在间硝基苯磺酸钠加氢反应中的应用" . | 工业催化 32 . 10 (2024) : 56-62 . |
| APA | 林知滨 , 郑辉东 , 范立海 , 吴丹 . Pd/C催化剂在间硝基苯磺酸钠加氢反应中的应用 . | 工业催化 , 2024 , 32 (10) , 56-62 . |
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D-allulose, a naturally occurring monosaccharide, is present in small quantities in nature. It is considered a valuable low-calorie sweetener due to its low absorption in the digestive tract and zero energy for growth. Most of the recent efforts to produce D-allulose have focused on in vitro enzyme catalysis. However, microbial fermentation is emerging as a promising alternative that offers the advantage of combining enzyme manufacturing and product synthesis within a single bioreactor. Here, a novel approach was proposed for the efficient biosynthesis of D-allulose from glycerol using metabolically engineered Escherichia coli. FbaA, Fbp, AlsE, and A6PP were used to construct the D-allulose synthesis pathway. Subsequently, PfkA, PfkB, and Pgi were disrupted to block the entry of the intermediate fructose-6-phosphate (F6P) into the Embden-Meyerhof-Parnas (EMP) and pentose phosphate (PP) pathways. Additionally, GalE and FryA were inactivated to reduce D-allulose consumption by the cells. Finally, a fed-batch fermentation process was implemented to optimize the performance of the cell factory. As a result, the titer of D-allulose reached 7.02 g/L with a maximum yield of 0.287 g/g.
Keyword ：
D-allulose D-allulose Escherichia coli Escherichia coli Fermentation Fermentation Metabolic engineering Metabolic engineering
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| GB/T 7714 | Guo, Qiang , Dong, Zhen-Xing , Luo, Xuan et al. Engineering Escherichia coli for D-allulose biosynthesis from glycerol [J]. | JOURNAL OF BIOTECHNOLOGY , 2024 , 394 : 103-111 . |
| MLA | Guo, Qiang et al. "Engineering Escherichia coli for D-allulose biosynthesis from glycerol" . | JOURNAL OF BIOTECHNOLOGY 394 (2024) : 103-111 . |
| APA | Guo, Qiang , Dong, Zhen-Xing , Luo, Xuan , Zheng, Ling-Jie , Fan, Li-Hai , Zheng, Hui-Dong . Engineering Escherichia coli for D-allulose biosynthesis from glycerol . | JOURNAL OF BIOTECHNOLOGY , 2024 , 394 , 103-111 . |
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Abstract ：
Sucrose is a commonly utilized nutritive sweetener in food and beverages due to its abundance in nature and low production costs. However, excessive intake of sucrose increases the risk of metabolic disorders, including diabetes and obesity. Therefore, there is a growing demand for the development of nonnutritive sweeteners with almost no calories. d-Allulose is an ultra-low-calorie, rare six-carbon monosaccharide with high sweetness, making it an ideal alternative to sucrose. In this study, we developed a cell factory for d-allulose production from sucrose using Escherichia coli JM109 (DE3) as a chassis host. The genes cscA, cscB, cscK, alsE, and a6PP were co-expressed for the construction of the synthesis pathway. Then, the introduction of ptsG-F and knockout of ptsG, fruA, ptsI, and ptsH to reprogram sugar transport pathways resulted in an improvement in substrate utilization. Next, the carbon fluxes of the Embden-Meyerhof-Parnas and the pentose phosphate pathways were regulated by the inactivation of pfkA and zwf, achieving an increase in d-allulose titer and yield of 154.2% and 161.1%, respectively. Finally, scaled-up fermentation was performed in a 5 L fermenter. The titer of d-allulose reached 11.15 g/L, with a yield of 0.208 g/g on sucrose.
Keyword ：
D-allulose D-allulose Escherichia coli Escherichia coli fermentation fermentation metabolic engineering metabolic engineering sucrose sucrose
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| GB/T 7714 | Zheng, Ling-Jie , Chen, Wei-Xiang , Zheng, Shang-He et al. Biosynthesis of nonnutritive monosaccharide d-allulose by metabolically engineered Escherichia coli from nutritive disaccharide sucrose [J]. | BIOTECHNOLOGY AND BIOENGINEERING , 2024 , 121 (12) : 3684-3693 . |
| MLA | Zheng, Ling-Jie et al. "Biosynthesis of nonnutritive monosaccharide d-allulose by metabolically engineered Escherichia coli from nutritive disaccharide sucrose" . | BIOTECHNOLOGY AND BIOENGINEERING 121 . 12 (2024) : 3684-3693 . |
| APA | Zheng, Ling-Jie , Chen, Wei-Xiang , Zheng, Shang-He , Ullah, Irfan , Zheng, Hui-Dong , Fan, Li-Hai et al. Biosynthesis of nonnutritive monosaccharide d-allulose by metabolically engineered Escherichia coli from nutritive disaccharide sucrose . | BIOTECHNOLOGY AND BIOENGINEERING , 2024 , 121 (12) , 3684-3693 . |
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D-Allulose, a C-3 epimer of d-fructose, has great market potential in food, healthcare, and medicine due to its excellent biochemical and physiological properties. Microbial fermentation for d-allulose production is being developed, which contributes to cost savings and environmental protection. A novel metabolic pathway for the biosynthesis of d-allulose from a d-xylose-methanol mixture has shown potential for industrial application. In this study, an artificial antisense RNA (asRNA) was introduced into engineered Escherichia coli to diminish the flow of pentose phosphate (PP) pathway, while the UDP-glucose-4-epimerase (GalE) was knocked out to prevent the synthesis of byproducts. As a result, the d-allulose yield on d-xylose was increased by 35.1%. Then, we designed a d-xylose-sensitive translation control system to regulate the expression of the formaldehyde detoxification operon (FrmRAB), achieving self-inductive detoxification by cells. Finally, fed-batch fermentation was carried out to improve the productivity of the cell factory. The d-allulose titer reached 98.6 mM, with a yield of 0.615 mM/mM on d-xylose and a productivity of 0.969 mM/h.
Keyword ：
d-allulose d-allulose Escherichia coli Escherichia coli fermentation fermentation metabolic engineering metabolic engineering
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| GB/T 7714 | Guo, Qiang , Zheng, Ling-Jie , Zheng, Shang-He et al. Enhanced Biosynthesis of d-Allulose from a D-Xylose-Methanol Mixture and Its Self-Inductive Detoxification by Using Antisense RNAs in Escherichia coli [J]. | JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY , 2024 , 72 (26) : 14821-14829 . |
| MLA | Guo, Qiang et al. "Enhanced Biosynthesis of d-Allulose from a D-Xylose-Methanol Mixture and Its Self-Inductive Detoxification by Using Antisense RNAs in Escherichia coli" . | JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY 72 . 26 (2024) : 14821-14829 . |
| APA | Guo, Qiang , Zheng, Ling-Jie , Zheng, Shang-He , Zheng, Hui-Dong , Lin, Xiao-Cheng , Fan, Li-Hai . Enhanced Biosynthesis of d-Allulose from a D-Xylose-Methanol Mixture and Its Self-Inductive Detoxification by Using Antisense RNAs in Escherichia coli . | JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY , 2024 , 72 (26) , 14821-14829 . |
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Fumarate is a value-added chemical that is widely used in food, medicine, material, and agriculture industries. With the rising attention to the demand for fumarate and sustainable development, many novel alternative ways that can replace the traditional petrochemical routes emerged. The in vitro cell-free multi-enzyme catalysis is an effective method to produce high value chemicals. In this study, a multi-enzyme catalytic pathway comprising three enzymes for fumarate production from low-cost substrates acetate and glyoxylate was designed. The acetylCoA synthase, malate synthase, and fumarase from Escherichia coli were selected and the coenzyme A achieved recyclable. The enzymatic properties and optimization of reaction system were investigated, reaching a fumarate yield of 0.34 mM with a conversion rate of 34% after 20 h of reaction. We proposed and realized the conversion of acetate and glyoxylate to fumarate in vitro using a cell-free multi-enzyme catalytic system, thus providing an alternative approach for the production of fumarate.
Keyword ：
Acetate Acetate Cell-free Cell-free Fumarate Fumarate Glyoxylate Glyoxylate Multi-enzyme catalysis Multi-enzyme catalysis
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| GB/T 7714 | Hou, Congli , Tian, Linyue , Lian, Guoli et al. Conversion of acetate and glyoxylate to fumarate by a cell-free synthetic enzymatic biosystem [J]. | SYNTHETIC AND SYSTEMS BIOTECHNOLOGY , 2023 , 8 (2) : 235-241 . |
| MLA | Hou, Congli et al. "Conversion of acetate and glyoxylate to fumarate by a cell-free synthetic enzymatic biosystem" . | SYNTHETIC AND SYSTEMS BIOTECHNOLOGY 8 . 2 (2023) : 235-241 . |
| APA | Hou, Congli , Tian, Linyue , Lian, Guoli , Fan, Li-Hai , Li, Zheng-Jun . Conversion of acetate and glyoxylate to fumarate by a cell-free synthetic enzymatic biosystem . | SYNTHETIC AND SYSTEMS BIOTECHNOLOGY , 2023 , 8 (2) , 235-241 . |
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Vibrio alginolyticus LHF01 was engineered to efficiently produce poly-3-hydroxybutyrate (PHB) from starch in this study. Firstly, the ability of Vibrio alginolyticus LHF01 to directly accumulate PHB using soluble starch as the carbon source was explored, and the highest PHB titer of 2.06 g/L was obtained in 18 h shake flask cultivation. Then, with the analysis of genomic information of V. alginolyticus LHF01, the PHB synthesis operon and amylase genes were identified. Subsequently, the effects of overexpressing PHB synthesis operon and amylase on PHB production were studied. Especially, with the co-expression of PHB synthesis operon and amylase, the starch consumption rate was improved and the PHB titer was more than doubled. The addition of 20 g/L insoluble corn starch could be exhausted in 6-7 h cultivation, and the PHB titer was 4.32 g/L. To the best of our knowledge, V. alginolyticus was firstly engineered to produce PHB with the direct utilization of starch, and this stain can be considered as a novel host to produce PHB using starch as the raw material.
Keyword ：
amylase amylase metabolic engineering metabolic engineering poly-3-hydroxybutyrate poly-3-hydroxybutyrate starch starch Vibrio alginolyticus Vibrio alginolyticus
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| GB/T 7714 | Li, Hong-Fei , Tian, Linyue , Lian, Guoli et al. Engineering Vibrio alginolyticus as a novel chassis for PHB production from starch [J]. | FRONTIERS IN BIOENGINEERING AND BIOTECHNOLOGY , 2023 , 11 . |
| MLA | Li, Hong-Fei et al. "Engineering Vibrio alginolyticus as a novel chassis for PHB production from starch" . | FRONTIERS IN BIOENGINEERING AND BIOTECHNOLOGY 11 (2023) . |
| APA | Li, Hong-Fei , Tian, Linyue , Lian, Guoli , Fan, Li-Hai , Li, Zheng-Jun . Engineering Vibrio alginolyticus as a novel chassis for PHB production from starch . | FRONTIERS IN BIOENGINEERING AND BIOTECHNOLOGY , 2023 , 11 . |
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