{"id":868,"date":"2024-11-20T07:37:15","date_gmt":"2024-11-20T07:37:15","guid":{"rendered":"https:\/\/www.biosynsis.org\/blog\/?p=868"},"modified":"2024-11-20T07:37:15","modified_gmt":"2024-11-20T07:37:15","slug":"insights-from-literature-mannheimia-succiniciproducens-metabolic-engineering-for-succinic-acid-production","status":"publish","type":"post","link":"https:\/\/www.biosynsis.com\/blog\/insights-from-literature-mannheimia-succiniciproducens-metabolic-engineering-for-succinic-acid-production\/","title":{"rendered":"Insights from Literature: Mannheimia succiniciproducens metabolic engineering for succinic acid production"},"content":{"rendered":"<div class=\"target-text-list\"><img decoding=\"async\" loading=\"lazy\" class=\"aligncenter size-full wp-image-869\" src=\"http:\/\/www.biosynsis.org\/blog\/wp-content\/uploads\/sites\/4\/2024\/11\/4.1-Mannheimia-succiniciproducens-metabolic-engineering-for-succinic-acid-production.png\" alt=\"Mannheimia succiniciproducens metabolic engineering for succinic acid production\" width=\"776\" height=\"401\" srcset=\"https:\/\/www.biosynsis.com\/blog\/wp-content\/uploads\/sites\/4\/2024\/11\/4.1-Mannheimia-succiniciproducens-metabolic-engineering-for-succinic-acid-production.png 776w, https:\/\/www.biosynsis.com\/blog\/wp-content\/uploads\/sites\/4\/2024\/11\/4.1-Mannheimia-succiniciproducens-metabolic-engineering-for-succinic-acid-production-300x155.png 300w, https:\/\/www.biosynsis.com\/blog\/wp-content\/uploads\/sites\/4\/2024\/11\/4.1-Mannheimia-succiniciproducens-metabolic-engineering-for-succinic-acid-production-768x397.png 768w\" sizes=\"(max-width: 776px) 100vw, 776px\" \/><\/div>\n<div class=\"target-text-list\">\n<div class=\"lg:flex lg:flex-col px-4 py-4\">\n<div class=\"sticky bottom-0 flex justify-between items-center px-3 py-2 bg-white dark:bg-dark-primary\">\n<div class=\"flex space-x-2.5 items-center\"><\/div>\n<\/div>\n<\/div>\n<div class=\"lg:flex lg:flex-col relative px-4 py-4\">\n<div id=\"mappedOutput\" class=\"transition bg-transparent !border-none p-0 mb-[3rem] h-full min-h-[30rem] !w-full text-black !ring-transparent focus:outline-none\">\n<p><span class=\"hover:bg-blue-100 dark:hover:bg-blue-400\/30 hover:cursor-pointer text-black dark:text-gray-200\" aria-expanded=\"false\">Succinic acid is a 4-carbon dicarboxylic acid.\u00a0<\/span><span class=\"hover:bg-blue-100 dark:hover:bg-blue-400\/30 hover:cursor-pointer text-black dark:text-gray-200\" aria-expanded=\"false\">Not only a major intermediate in the TCA, it is also a major raw material in food, pharmaceutical and chemical production.\u00a0<\/span><span class=\"hover:bg-blue-100 dark:hover:bg-blue-400\/30 hover:cursor-pointer text-black dark:text-gray-200\" aria-expanded=\"false\">Since the market for sustainable development is increasing, renewable energy for microbial fermentation of succinic acid is gaining traction in the industry.\u00a0<\/span><span class=\"hover:bg-blue-100 dark:hover:bg-blue-400\/30 hover:cursor-pointer text-black dark:text-gray-200\" aria-expanded=\"false\">But old-school fermentation tends to be coupled with the production of byproducts (acetic acid, formic acid, lactic acid) that diminish yields and cost extra to purify.<\/span><\/p>\n<p><span class=\"hover:bg-blue-100 dark:hover:bg-blue-400\/30 hover:cursor-pointer text-black dark:text-gray-200\" aria-expanded=\"false\">The team\u2019s solution to this challenge was to modify the gram-negative anaerobic bacteria Mannheimia succiniciproducens through genomics and metabolic engineering, and construct a highly productive strain of succinic acid producing bacteria that reduced by-products significantly.\u00a0<\/span><span class=\"hover:bg-blue-100 dark:hover:bg-blue-400\/30 hover:cursor-pointer text-black dark:text-gray-200\" aria-expanded=\"false\">These results demonstrate the power of genome-scale metabolic engineering to optimise biobased chemical manufacturing.<\/span><\/p>\n<\/div>\n<div><img decoding=\"async\" loading=\"lazy\" class=\"aligncenter size-full wp-image-871\" src=\"http:\/\/www.biosynsis.org\/blog\/wp-content\/uploads\/sites\/4\/2024\/11\/4.2-Succinic-Acid-biosynthesis-pathway.png\" alt=\"Succinic Acid biosynthesis pathway\" width=\"835\" height=\"574\" srcset=\"https:\/\/www.biosynsis.com\/blog\/wp-content\/uploads\/sites\/4\/2024\/11\/4.2-Succinic-Acid-biosynthesis-pathway.png 835w, https:\/\/www.biosynsis.com\/blog\/wp-content\/uploads\/sites\/4\/2024\/11\/4.2-Succinic-Acid-biosynthesis-pathway-300x206.png 300w, https:\/\/www.biosynsis.com\/blog\/wp-content\/uploads\/sites\/4\/2024\/11\/4.2-Succinic-Acid-biosynthesis-pathway-768x528.png 768w\" sizes=\"(max-width: 835px) 100vw, 835px\" \/><\/div>\n<div style=\"text-align: center;\"><em>Succinic Acid biosynthesis pathway<\/em><\/div>\n<div><\/div>\n<\/div>\n<\/div>\n<h2 class=\"target-text-list\">Overview<\/h2>\n<div class=\"target-text-list\">This study aims to optimize M. Succiniciproducens metabolic pathways to:<\/div>\n<ul>\n<li class=\"target-text-list\">Increase succinic acid production;<\/li>\n<li class=\"target-text-list\">Eliminate the formation of by-products during the fermentation process;<\/li>\n<li class=\"target-text-list\">Explore the role of key metabolic enzymes in succinate synthesis.<\/li>\n<\/ul>\n<div><\/div>\n<h2 class=\"target-text-list\">Research methods and strategies<\/h2>\n<div><\/div>\n<p><strong>1 Gene knockout strategy:<\/strong><\/p>\n<div class=\"target-text-list\"><\/div>\n<ul>\n<li class=\"target-text-list\">Carry out gradual knock-out of key enzyme genes for by-product production (such as ldhA, pflB, pta and ackA).<\/li>\n<li class=\"target-text-list\">Experiments were conducted to verify the impact of gene knockout on metabolic flow redistribution and succinate production.<\/li>\n<\/ul>\n<div><img decoding=\"async\" loading=\"lazy\" class=\"aligncenter size-full wp-image-872\" src=\"http:\/\/www.biosynsis.org\/blog\/wp-content\/uploads\/sites\/4\/2024\/11\/4.3-Strategies-for-disrupting-the-ldhA-and-pflB-genes-in-the-chromosome-of-M.-succiniproducens.png\" alt=\"Strategies for disrupting the ldhA and pflB genes in the chromosome of M. succiniproducens\" width=\"726\" height=\"825\" srcset=\"https:\/\/www.biosynsis.com\/blog\/wp-content\/uploads\/sites\/4\/2024\/11\/4.3-Strategies-for-disrupting-the-ldhA-and-pflB-genes-in-the-chromosome-of-M.-succiniproducens.png 726w, https:\/\/www.biosynsis.com\/blog\/wp-content\/uploads\/sites\/4\/2024\/11\/4.3-Strategies-for-disrupting-the-ldhA-and-pflB-genes-in-the-chromosome-of-M.-succiniproducens-264x300.png 264w\" sizes=\"(max-width: 726px) 100vw, 726px\" \/><\/div>\n<div style=\"text-align: center;\"><em>Strategies for disrupting the ldhA and pflB genes in the chromosome of M. succiniproducens<\/em><\/div>\n<div><\/div>\n<p><strong>2 Fermentation process optimization:<\/strong><\/p>\n<div class=\"target-text-list\"><\/div>\n<ul>\n<li class=\"target-text-list\">Compare succinate production under batch culture and fed-batch culture conditions;<\/li>\n<li class=\"target-text-list\">Use optimized medium compositions and CO \u02c7 supply to enhance fermentation efficiency.<\/li>\n<\/ul>\n<div><\/div>\n<p><strong>3 Analysis of key metabolic enzymes:<\/strong><\/p>\n<div class=\"target-text-list\"><\/div>\n<ul>\n<li class=\"target-text-list\">To study the role of three CO \u02c7-fixing enzymes (PEP carboxylase, PEP carboxykinase and malase) in succinate synthesis and analyze their contribution to the metabolic flux of C-C.<\/li>\n<\/ul>\n<div>\n<div class=\"info-background\">\n<h2>Related Services<\/h2>\n<p><a href=\"\/metabolic-engineering-and-pathway-design.html\">Metabolic Engineering and Pathway Design<\/a><\/p>\n<p><a href=\"\/bacterial-metabolic-engineering-services.html\">Bacterial Metabolic Engineering<\/a><\/p>\n<p><a href=\"\/crispr-based-gene-editing-services.html\">CRISPR-based gene editing services<\/a><\/p>\n<p><a href=\"\/crispr-library-construction-service.html\">CRISPR Library Construction Service<\/a><\/p>\n<p><a href=\"\/saccharomyces-cerevisiae-metabolic-engineering-services.html\">Saccharomyces cerevisiae Metabolic Engineering Services<\/a><\/p>\n<p><a href=\"\/succinic-acid-biosynthesis-development-service.html\">Strain Engineering for Succinic Acid<\/a><\/p>\n<\/div>\n<\/div>\n<h2 class=\"target-text-list\"><span class=\"text-black dark:text-gray-200\">Key findings<\/span><\/h2>\n<p><strong><b>1.Optimization of metabolic pathways and its effects<\/b><\/strong><\/p>\n<p>The gene knockout approach was successful as the research group had transformed M. Metabolic network of succiniciproducens:<\/p>\n<ul>\n<li>Inhibition of lactate production:<\/li>\n<\/ul>\n<p>After knocking out the gene for lactate dehydrogenase (ldhA), lactic acid production fell sharply, succinic acid was produced at a higher rate (10.5 g\/L to 12.0 g\/L), and the conversion rate was 0.90 mol\/mol glucose.<\/p>\n<p>&nbsp;<\/p>\n<ul>\n<li>Decrease formic acid and acetic acid formation:<\/li>\n<\/ul>\n<p>&nbsp;<\/p>\n<p>After knocking out ldhA, the pflB (pyruvate formate lyase) gene was also knocked out and formic acid was produced at 100% without the presence of any acetate.\u00a0Now the succinic acid yield was 13.4 g\/L and the conversion was 0.97 mol\/mol glucose.<\/p>\n<p>&nbsp;<\/p>\n<ul>\n<li>Whole-body blockage of acetic acid formation:<\/li>\n<\/ul>\n<p>Once the genes for pta (phosphotransacetylase) and ackA (acetate kinase) were deleted, acetate production was practically eliminated.\u00a0The final succinic acid production of the modified strain (LPK7) was 13.4 g\/L, with acetate and lactic acid almost invisible.<\/p>\n<p>&nbsp;<\/p>\n<p>&nbsp;<\/p>\n<ol start=\"2\">\n<li><strong><b>Contribution of CO fixation mechanisms<\/b><\/strong><\/li>\n<\/ol>\n<p>The three CO -fixing enzymes that were investigated each contribute different components to succinate synthesis:<\/p>\n<p>&nbsp;<\/p>\n<ul>\n<li>PEP carboxykinase (pckA):<\/li>\n<\/ul>\n<p>It was discovered to be an important enzyme in C to C metabolic flux conversion.\u00a0Its knockout drastically lowers the succinate (by 57%) and slows the strain\u2019s anaerobic development.<\/p>\n<p>&nbsp;<\/p>\n<ul>\n<li>PEP carboxylase (ppc) and malylase (maeB):<\/li>\n<\/ul>\n<p>Its knockout is less detrimental to succinic acid synthesis, but alters the distribution pattern of by-products.<\/p>\n<p>&nbsp;<\/p>\n<ol start=\"3\">\n<li><strong><b>Significant advantages of fed-batch fermentation<\/b><\/strong><\/li>\n<\/ol>\n<p>The succinic acid production of LPK7 strain was further augmented to 52.4 g\/L under fed-batch fermentation, and the productivity increased to 1.8 g\/L\/h.\u00a0Experiments also showed that:<\/p>\n<p>&nbsp;<\/p>\n<ul>\n<li>By-products were not produced at all, and there was only a trace of malic acid and pyruvate;<\/li>\n<li>It climbed to 1.16 mol\/mol glucose (0.76 g\/g glucose), which is among the highest seen.<\/li>\n<\/ul>\n<div><\/div>\n<h2 class=\"target-text-list\">Industrialization potential and challenges<\/h2>\n<div class=\"target-text-list\">\n<p><strong><b>T<\/b><\/strong><strong><b>echnical advantages<\/b><\/strong><\/p>\n<ol>\n<li>Efficient by-product inhibition:<\/li>\n<\/ol>\n<p>Compared with other fermentation strains, the succinic acid production of LPK7 strain is hardly accompanied by the production of acetic acid, formic acid and lactic acid, greatly simplifying the downstream purification process.<\/p>\n<ol start=\"2\">\n<li>Sustainability:<\/li>\n<\/ol>\n<p>Using carbon dioxide as a reaction substrate not only increases succinic acid production, but also achieves effective recycling of carbon resources.<\/p>\n<ol start=\"3\">\n<li>High conversion and production rates:<\/li>\n<\/ol>\n<p>The optimized fermentation process can significantly reduce raw material consumption and production costs, providing feasibility for large-scale industrial production.<\/p>\n<p>&nbsp;<\/p>\n<p><strong><b>E<\/b><\/strong><strong><b>xisting challenges<\/b><\/strong><\/p>\n<ol>\n<li>Pyruvic acid and malic acid residues:<\/li>\n<\/ol>\n<p>Although major by-products were suppressed, the accumulation of pyruvate and malate limited further improvements in succinic acid production.<\/p>\n<ol start=\"2\">\n<li>Insufficient supply of reducing power:<\/li>\n<\/ol>\n<p>The production of succinic acid requires a large amount of reducing power, and in the future, consideration can be given to enhancing electron supply through external hydrogen supply or optimizing the expression of reducing power-related enzymes.<\/p>\n<ol start=\"3\">\n<li>Genome stability:<\/li>\n<\/ol>\n<p>The genetic stability of engineered strains in long-term fermentation or industrial applications requires further verification.<\/p>\n<\/div>\n","protected":false},"excerpt":{"rendered":"<p>Succinic acid is a 4-carbon dicarboxylic acid.\u00a0Not only a major intermediate in the TCA, it is also a major raw material in food, pharmaceutical and chemical production.\u00a0Since the market for sustainable development is increasing, renewable energy for microbial fermentation of succinic acid is gaining traction in the industry.\u00a0But old-school fermentation tends to be coupled with&#8230;<\/p>\n","protected":false},"author":1,"featured_media":870,"comment_status":"closed","ping_status":"closed","sticky":false,"template":"","format":"standard","meta":[],"categories":[1],"tags":[],"aioseo_notices":[],"_links":{"self":[{"href":"https:\/\/www.biosynsis.com\/blog\/wp-json\/wp\/v2\/posts\/868"}],"collection":[{"href":"https:\/\/www.biosynsis.com\/blog\/wp-json\/wp\/v2\/posts"}],"about":[{"href":"https:\/\/www.biosynsis.com\/blog\/wp-json\/wp\/v2\/types\/post"}],"author":[{"embeddable":true,"href":"https:\/\/www.biosynsis.com\/blog\/wp-json\/wp\/v2\/users\/1"}],"replies":[{"embeddable":true,"href":"https:\/\/www.biosynsis.com\/blog\/wp-json\/wp\/v2\/comments?post=868"}],"version-history":[{"count":109,"href":"https:\/\/www.biosynsis.com\/blog\/wp-json\/wp\/v2\/posts\/868\/revisions"}],"predecessor-version":[{"id":873,"href":"https:\/\/www.biosynsis.com\/blog\/wp-json\/wp\/v2\/posts\/868\/revisions\/873"}],"wp:featuredmedia":[{"embeddable":true,"href":"https:\/\/www.biosynsis.com\/blog\/wp-json\/wp\/v2\/media\/870"}],"wp:attachment":[{"href":"https:\/\/www.biosynsis.com\/blog\/wp-json\/wp\/v2\/media?parent=868"}],"wp:term":[{"taxonomy":"category","embeddable":true,"href":"https:\/\/www.biosynsis.com\/blog\/wp-json\/wp\/v2\/categories?post=868"},{"taxonomy":"post_tag","embeddable":true,"href":"https:\/\/www.biosynsis.com\/blog\/wp-json\/wp\/v2\/tags?post=868"}],"curies":[{"name":"wp","href":"https:\/\/api.w.org\/{rel}","templated":true}]}}