Heterologous biosynthesis of tetrahydrocannabinolic acid (THCA) in yeast is a biotechnological process in Natural Product Biotechnology that was recently introduced. Based on heterologous genes from Cannabis sativa and Streptomyces spp. cloned into Saccharomyces cerevisiae, the heterologous biosynthesis was fully embedded as a proof of concept. Low titer and insufficient biocatalytic rate of most enzymes require systematic optimization of recombinant catalyst by protein engineering and consequent C-flux improvement of the yeast chassis for sufficient precursor (acetyl-CoA), energy (ATP), and NADH delivery. In this review basic principles of in silico analysis of anabolic pathways towards olivetolic acid (OA) and cannabigerolic acid (CBGA) are elucidated and discussed to identify metabolic bottlenecks. Based on own experimental results, yeasts are discussed as potential platform organisms to be introduced as potential cannabinoid biofactories. Especially feeding strategies and limitations in the committed mevalonate and olivetolic acid pathways are in focus of in silico and experimental studies to validate the scientific and commercial potential as a realistic alternative to the plant Cannabis sativa.

Key points

• First time critical review of the heterologous process for recombinant THCA/CBDA production and critical review of bottlenecks and limitations for a bioengineered technical process

• Integrative approach of protein engineering, systems biotechnology, and biochemistry of yeast physiology and biosynthetic cannabinoid enzymes

• Comparison of NphB and CsPT aromatic prenyltransferases as rate-limiting catalytic steps towards cannabinoids in yeast as platform organisms

酵母中四氢大麻酚酸（THCA）的异源生物合成是最近引入的天然产物生物技术中的生物技术过程。基于来自大麻和链霉菌属物种的异源基因。克隆到酿酒酵母中，异源生物合成被完全嵌入作为概念证明。大多数酶的滴度低且生物催化速率不足，需要通过蛋白质工程系统地优化重组催化剂，并相应地改善酵母底盘的C-通量，以提供足够的前体（乙酰-CoA），能量（ATP）和NADH。在这篇综述中，阐明并讨论了合成代谢途径向橄榄酸（OA）和大麻二酸（CBGA）的计算机模拟分析的基本原理，以识别代谢瓶颈。根据自己的实验结果，将酵母作为潜在的平台生物进行讨论，以作为潜在的大麻素生物工厂引入。大麻。

关键点

•首次对重组THCA / CBDA生产的异源过程进行严格审查，并对生物工程技术过程的瓶颈和局限性进行严格审查

•蛋白质工程，系统生物技术以及酵母生理学和生物合成大麻素酶生物化学的综合方法

•比较NphB和CsPT芳香异戊二烯基转移酶作为对作为平台生物的酵母中大麻素的限速催化步骤