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Chinese Journal of Biotechnology ; (12): 1721-1736, 2021.
Article in Chinese | WPRIM | ID: wpr-878663


Higher alcohols that contain more than two carbon atoms have better fuel properties than ethanol, making them important supplements and alternatives to fossil fuels. Using microbes to produce higher alcohols from renewable biomass can alleviate the current energy and environmental crises, and has become a major future direction for green biomanufacturing. Since natural microbes can only produce a few higher alcohols in small amounts, it is necessary to reconstruct the synthetic pathways for higher alcohols in model industrial strains through metabolic engineering and synthetic biology to overcome the metabolic bottlenecks. A series of milestones have been accomplished in past decades. The authors of this review have witnessed the entire journey of this field from its first success to the leaping development. On the 30th anniversary of the founding of the discipline of metabolic engineering, this review dates back to the great milestones in achieving heterologous production of higher alcohols in non-native strains. The design and optimization of high alcohol biosynthetic pathways, the expansion of feedstock, the engineering of host strains and the industrialization process are summarized. This review aims to draw further attention to microbial synthesis of higher alcohols, inspire the development of novel techniques and strategies of metabolic engineering, and promote the innovation and upgrade of China's biofuel industry.

Alcohols , Biofuels , Biosynthetic Pathways , Ethanol , Metabolic Engineering , Synthetic Biology
Chinese Journal of Biotechnology ; (12): 429-447, 2021.
Article in Chinese | WPRIM | ID: wpr-878573


Higher alcohols are one of the main by-products of Saccharomyces cerevisiae in brewing. High concentration of higher alcohols in alcoholic beverages easily causes headache, thirst and other symptoms after drinking. It is also the main reason for chronic drunkenness and difficulty in sobering up after intoxication. The main objective of this review is to present an overview of the flavor characteristics and metabolic pathways of higher alcohols as well as the application of mutagenesis breeding techniques in the regulation of higher alcohol metabolism in S. cerevisiae. In particular, we review the application of metabolic engineering technology in genetic modification of amino transferase, α-keto acid metabolism, acetate metabolism and carbon-nitrogen metabolism. Moreover, key challenges and future perspectives of realizing optimization of higher alcohols metabolism are discussed. This review is intended to provide a comprehensive understanding of metabolic regulation system of higher alcohols in S. cerevisiae and to provide insights into the rational development of the excellent industrial S. cerevisiae strains producing higher alcohols.

Alcoholic Beverages , Alcohols/analysis , Fermentation , Saccharomyces cerevisiae/metabolism , Saccharomyces cerevisiae Proteins/metabolism
Chinese Journal of Biotechnology ; (12): 522-534, 2019.
Article in Chinese | WPRIM | ID: wpr-771355


As a new beer fermentation technology, high temperature and high gravity fermentation has brought many benefits to brewery industry, but there are also a series of problems such as the decrease of yeast flocculation ability at the end of fermentation and the high concentration of higher alcohols. To increase yeast flocculation ability and reduce the production of higher alcohols in high temperature and high gravity fermentation of beer, BAT2 was replaced by the FLO5 expression cassette to obtain the mutant strain S6-BF2. Real-time quantitative PCR showed that the relative transcriptional level of FLO5 in S6-BF2 improved 17.8 times compared with that in S6. The flocculation ability of mutant S6-BF2 heightened by 63% compared to that of the original strain S6, and the concentration of higher alcohols decreased from 175.58 mg/L to 159.58 mg/L in high temperature and high gravity fermentation of beer. Moreover, the activity of mitochondrial branched-chain amino acid transferase was repressed, resulting in the production of higher alcohols of 142.13 mg/L, reduced by 18.4% compared to that of the original strain S6, meanwhile, the flocculation ability of mutant S6-BF2B1 kept unchanged compared to the mutant S6-BF2. The determination result of flavor compounds showed that the higher alcohols/ester ratio in beer was reasonable. This research has suggested an effective strategy for enhancing yeast flocculation ability and decreasing production of higher alcohols in high-temperature and high-gravity brewing.

Beer , Fermentation , Hypergravity , Saccharomyces cerevisiae , Saccharomyces cerevisiae Proteins , Temperature , Transaminases
Acta biol. colomb ; 17(2): 309-322, mayo-ago. 2012. ilus, graf, tab
Article in Spanish | LILACS-Express | LILACS | ID: lil-659299


Se ha estudiado la influencia de la aireación en la actividad fermentativa de Kloeckera apiculata RIVE 9-2-1 con el objetivo de evaluar la producción de metabolitos de la fermentación. Para ello, la cepa se cultivó en frascos Erlenmeyer conteniendo jugo de manzana esterilizado y sin aroma, y los compuestos químicos producidos durante la fermentación en cultivo agitado (200 min-1) y estático (sin agitación) se determinaron. Los resultados mostraron que la agitación del medio de cultivo incrementa la producción de alcoholes superiores (hasta 591,0 mg/L) comparado al cultivo estático, mientras que por el contrario, la producción de ácido acético, etil acetato y glicerol (260,0 ± 11,0 mg/L, 196,0 ± 10,0 mg/L y 2,6 ± 0,2 g/L) resultaron ser mayores que en cultivo agitado (222,0 ± 8,0 mg/L, 96,0 ± 4,5 mg/L y 1,8 ± 0,2 g/L) respectivamente. Cultivos bacth realizados en biorreactor con un flujo de aire de 25 L/h reportaron una tasa de crecimiento (µ) de 0,17 h-1, producción de etanol (12,5 ± 2,0 g/L) y otros compuestos típicamente producidos durante la fermentación alcohólica. La concentración de oxígeno disuelto en el medio de fermentación afecta su metabolismo así, cantidades insuficientes de oxígeno provocaría un metabolismo respirofermentativo. Los mejores resultados en términos de calidad organoléptica de la bebida fermentada en lo referente al aroma, sabor y olor se obtuvieron en la fermentación en cultivo estático. El control de la aireación del medio de fermentación puede ser usado para controlar la síntesis de compuestos químicos de impacto sensorial en la producción de bebidas fermentadas.

The influence of aireation on the fermentative activity of Kloeckera apiculata RIVE 9-2-1 was studied in order to evaluate the production of metabolites of the fermentation. To achieve this, the strain was cultured in Erlenmeyer flasks containing sterilized and aroma removed apple juice, and the chemical compounds produced during fermentation in shaken (200 min-1) and static (without agitation) cultivation were determined. The results showed that the agitation of the culture medium increases production of higher alcohols (till 591.0 mg/L) compared to static cultivation, whereas on the contrary, the production of acetic acid, ethyl acetate and glycerol (260.0 ± 11.0 mg/L, 196.0 ± 10.0 mg/L y 2.6±0.2 g/L) were higher compared to shaken cultivation (222.0 ± 8.0 mg/L, 96.0 ± 4.5 mg/L and 1.8 ± 0.2 g/L) respectively. Batch cultivations carried out in bioreactor with air flux of 25 l/h reported a growth rate (µ) of 0.17 h-1, production of ethanol (12.5 ± 2.0 g/L) and other compounds typically produced during alcoholic fermentation. The concentration of dissolved oxygen in the fermentation medium affects its metabolism thus; insufficient amounts of oxygen would provoke a respirofermentative metabolism. The best results in terms of organoleptic quality of the fermented beverage regarding to aroma, taste and flavour was obtained when fermented in static cultivation. The control of aeration during fermentation can be used to control the synthesis of chemical compounds of sensory impact in the production of fermented beverages.