An in Vitro Biosynthesis of Sesquiterpenes Starting from Acetic Acid

Dirkmann, Michael; Nowack, Julia; Schulz, Frank

doi:10.1002/cbic.201800128

Cited by 22 publications

(28 citation statements)

References 29 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…In contrast, it was observed for a carbohydrate precursor synthesis using a four-enzyme cascade that a reduction of reaction temperature from 30 • C to 20 • C led to an increase in yield from 87% to above 95% [37]. The production of sesquiterpenes by Dirkmann et al is another example for the importance of the reaction temperature since they found 30 • C to be the optimal reaction temperature [47]. Temperatures below reduced the conversion and above led to protein precipitation.…”

Section: Reaction Temperaturementioning

confidence: 99%

“…Metabolic pathways already existing in organisms can be used as templates and transferred to in vitro multi-enzymatic reactions. These can be parts of the primary metabolism, e.g., glycolysis [46] or of the secondary metabolism such as the mevalonate pathway [47] or chitin degradation [35]. Combinations of primary and secondary pathways were developed to in vitro cascades as well, which produces high value products with the benefit of energy regeneration and the usage of bulk chemicals as substrates.…”

Section: Cascade Design Reaction Pathwaymentioning

confidence: 99%

See 1 more Smart Citation

Getting the Most Out of Enzyme Cascades: Strategies to Optimize In Vitro Multi-Enzymatic Reactions

et al. 2021

View full text Add to dashboard Cite

In vitro enzyme cascades possess great benefits, such as their synthetic capabilities for complex molecules, no need for intermediate isolation, and the shift of unfavorable equilibria towards the products. Their performance, however, can be impaired by, for example, destabilizing or inhibitory interactions between the cascade components or incongruous reaction conditions. The optimization of such systems is therefore often inevitable but not an easy task. Many parameters such as the design of the synthesis route, the choice of enzymes, reaction conditions, or process design can alter the performance of an in vitro enzymatic cascade. Many strategies to tackle this complex task exist, ranging from experimental to in silico approaches and combinations of both. This review collates examples of various optimization strategies and their success. The feasibility of optimization goals, the influence of certain parameters and the usage of algorithm-based optimizations are discussed.

show abstract

Section: Reaction Temperaturementioning

confidence: 99%

Section: Cascade Design Reaction Pathwaymentioning

confidence: 99%

Getting the Most Out of Enzyme Cascades: Strategies to Optimize In Vitro Multi-Enzymatic Reactions

et al. 2021

View full text Add to dashboard Cite

show abstract

“…The highest yields with more than 95% and titers of 15 g•L −1 were achieved with a cell-free system consisting of 27 purified enzymes, which convert glucose into monoterpenes [31]. Other systems which incorporate acetic acid as a starting building block for the cell-free synthesis of terpenes are described as well [32]. However, a major drawback is the need for purified enzymes, which are associated with additional costs and the further input of glucose needed to produce them.…”

Section: Progess To An Economic Limonene Production Processmentioning

confidence: 99%

A Gram-Scale Limonene Production Process with Engineered Escherichia coli

et al. 2020

View full text Add to dashboard Cite

Monoterpenes, such as the cyclic terpene limonene, are valuable and important natural products widely used in food, cosmetics, household chemicals, and pharmaceutical applications. The biotechnological production of limonene with microorganisms may complement traditional plant extraction methods. For this purpose, the bioprocess needs to be stable and ought to show high titers and space-time yields. In this study, a limonene production process was developed with metabolically engineered Escherichia coli at the bioreactor scale. Therefore, fed-batch fermentations in minimal medium and in the presence of a non-toxic organic phase were carried out with E. coli BL21 (DE3) pJBEI-6410 harboring the optimized genes for the mevalonate pathway and the limonene synthase from Mentha spicata on a single plasmid. The feasibility of glycerol as the sole carbon source for cell growth and limonene synthesis was examined, and it was applied in an optimized fermentation setup. Titers on a gram-scale of up to 7.3 g·Lorg−1 (corresponding to 3.6 g·L−1 in the aqueous production phase) were achieved with industrially viable space-time yields of 0.15 g·L−1·h−1. These are the highest monoterpene concentrations obtained with a microorganism to date, and these findings provide the basis for the development of an economic and industrially relevant bioprocess.

show abstract

“…However, Schi. pombe has mevalonate pathway to generate terpenes from acetic acid, thereby reducing the stress of acetic acid ultimately in microbiota [43,44]. Therefore, studies need to explore further information about the microbial interactions between S. cerevisiae and non-Saccharomyces yeasts at the molecular level.…”

Section: Discussionmentioning

confidence: 99%

Schizosaccharomyces pombe Can Reduce Acetic Acid Produced by Baijiu Spontaneous Fermentation Microbiota

Song

Zhang

et al. 2019

Microorganisms

View full text Add to dashboard Cite

The spontaneous fermentation of alcoholic beverage is a bioprocess donated by microbiota with complex stress environments. Among various microbes, non-Saccharomyces yeasts have high stress tolerance and significantly affect the taste and quality of products in process. Although many researchers have focused on the influence of acid stress, the mechanism of non-Saccharomyces yeasts to tolerant stress remains unclear in microbiota. To bridge the gap, we constructed in situ and in vitro studies to explore the reduction pathway of acetic acid in non-Saccharomyces yeasts. In this study, we found Schizosaccharomyces pombe has special capacities to resist 10 g/L acetic acid in laboratory cultures and decrease the average concentration of acetic acid from 9.62 to 6.55 g/kg fermented grains in Chinese Maotai-flavor liquor (Baijiu) production. Moreover, Schi. pombe promoted metabolic level of mevalonate pathway (high expressions of gene ACCAT1, HMGCS1, and HMGCR1) to degrade a high concentration of acetic acid. Meanwhile, Schi. pombe also improved the concentration of mevalonic acid that is the precursor of terpenes to enhance the taste and quality of Baijiu. Overall, the synchronicity of reduction and generation in Schi. pombe advances the current knowledge to guide more suitable strategies for mechanism studies of non-Saccharomyces yeasts in fermented industries of alcoholic beverages.

show abstract

An in Vitro Biosynthesis of Sesquiterpenes Starting from Acetic Acid

Cited by 22 publications

References 29 publications

Getting the Most Out of Enzyme Cascades: Strategies to Optimize In Vitro Multi-Enzymatic Reactions

Getting the Most Out of Enzyme Cascades: Strategies to Optimize In Vitro Multi-Enzymatic Reactions

A Gram-Scale Limonene Production Process with Engineered Escherichia coli

Schizosaccharomyces pombe Can Reduce Acetic Acid Produced by Baijiu Spontaneous Fermentation Microbiota

Contact Info

Product

Resources

About