转基因植物中酚类物质的生物漏斗表达细菌3-脱氢莽草酸脱水酶(qsuB)基因

IF 2.5 Q3 BIOTECHNOLOGY & APPLIED MICROBIOLOGY Frontiers in chemical engineering Pub Date : 2022-10-19 DOI:10.3389/fceng.2022.1036084
G. Umana, Jose M. Perez, Faride Unda, Chien-Yuan Lin, Canan Sener, S. D. Karlen, S. Mansfield, A. Eudes, J. Ralph, T. Donohue, D. Noguera
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引用次数: 3

摘要

木质纤维素生物质生物炼制的经济和环境可持续性取决于从细胞壁多糖和木质素聚合物中生产生物燃料和生物产品。植物基因工程的历史努力集中在开发促进生物质解构的策略上,最近的努力包括在植物中合成高价值的化学物质。一种这样的遗传修饰是细菌奎宁酸盐和莽草酸盐利用B(qsuB)基因的表达,其增加了原儿茶酸在木质纤维素生物质中的积累。在此,我们评估了碱性预处理工艺的有效性,该工艺可直接从拟南芥、白杨和高粱的野生型和QsuB转基因系中提取酚类物质,然后用芳构新鞘氨醇的工程菌株将其升级为聚酯前体2-吡咯-4,6-二羧酸(PDC)。从所有QsuB转基因品系中提取的原儿茶酸大多呈糖基化形式。糖基化原儿茶酸酯和其他植物衍生的酚类物质被芳香菌有效代谢,使用拟南芥QsuB基因转基因品系的提取物(~5%w/w)产生PDC最大,其次是QsuB高粱(~1.1%w/w),和QsuB杨树(~0.4%w/w)品系。拟南芥、白杨和高粱的野生型和QsuB转基因系的PDC生产的比较表明,温和的碱性预处理可以从植物生物质中释放酚类物质,这些植物生物质要么是天然存在的,要么是由于基因工程策略而积累的。就观察到的PDC产量而言,所有QsuB转基因品系都优于野生型对应品系。此外,即使当提取的大部分原儿茶酸是糖基化形式时,微生物向PDC的漏斗输送也是有效的,这清楚地表明这种细菌可以代谢这些芳香缀合物。这些发现说明了植物和微生物工程相结合在木质纤维素生物精炼厂中由酚类物质形成生物产品的好处。
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Biological funneling of phenolics from transgenic plants engineered to express the bacterial 3-dehydroshikimate dehydratase (qsuB) gene
The economic and environmental sustainability of lignocellulosic biomass biorefineries is predicated on generating biofuels and bioproducts from cell-wall polysaccharide and lignin polymers. Historical efforts in plant genetic engineering have focused on the development of strategies that facilitate biomass deconstruction, with more recently efforts including the synthesis of high-value chemicals in planta. One such genetic modification is the expression of the bacterial quinate and shikimate utilization B (qsuB) gene that increases the accumulation of protocatechuic acid in lignocellulosic biomass. Herein, we evaluated the effectiveness of an alkaline pretreatment process to extract phenolics directly from wild-type and QsuB-transgenic lines of Arabidopsis, poplar, and sorghum, and then upgrade them to the polyester precursor 2-pyrone-4,6-dicarboxylic acid (PDC) with an engineered strain of Novosphingobium aromaticivorans. Protocatechuic acid extracted from all QsuB transgenic lines was found to be mostly in the glycosylated form. Glycosylated protocatechuic acid and other plant-derived phenolics were effectively metabolized by N. aromaticivorans, and PDC production was greatest using extracts from an Arabidopsis QsuB transgenic line (∼5% w/w), followed by QsuB sorghum (∼1.1% w/w), and QsuB poplar (∼0.4% w/w) lines. The comparison of PDC production from wild-type and QsuB transgenic lines of Arabidopsis, poplar, and sorghum demonstrates the utility of a mild alkaline pretreatment to liberate phenolics from plant biomass that are either naturally present or that accumulate as a consequence of genetic engineering strategies. All QsuB transgenic lines outperformed their wild-type counterparts with respect to observed PDC yields. In addition, microbial funneling to PDC was effective even when most of the protocatechuic acid extracted was in glycosylated form, clearly demonstrating that this bacterium can metabolize these aromatic conjugates. These findings illustrate the benefits of combining plant and microbial engineering for bioproduct formation from phenolics in lignocellulosic biorefineries.
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