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Construction of crocin biosynthesis pathway in Saccharomyces cerevisiae
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Advisor
이평천
Affiliation
아주대학교 일반대학원
Department
일반대학원 분자과학기술학과
Publication Year
2022-02
Publisher
The Graduate School, Ajou University
Keyword
crocin
Description
학위논문(석사)--아주대학교 일반대학원 :분자과학기술학과,2022. 2
Alternative Abstract
Crocins belong to the apocarotenoid glycosides and are the main bioactive components in saffron, derived from the stigmas of Crocus sativus. Under the need for efficient and economical methods for producing crocins, microbial production enhances the possibility of fabrication of crocins. Four major heterologous enzymes were introduced in S.cerevisiae for simulating the crocin biosynthetic pathway. β-carotene hydroxylase (CrtZ), carotenoid cleavage dioxygenase (CCD), aldehyde dehydrogenase (ALDH), and two types of UDP-glucosyltransferases (UGTs) were expressed, including novel enzymes that were isolated by our group. A qualitative-quantitative analysis of the precursor was performed at each step to find the optimal catalytic activities among candidate enzymes from different organism sources. As a result, the metabolic engineered S.cerevisiae strain produced 65.23 ± 2.97μg/L of crocin-4 in flask cultivation. To increase the supply of UDP-glucose, two genes named pgm2, ugp1 involved in endogenous uridine diphosphate glucose pathway were over-expressed respectively in S. cerevisiae. As a result, overexpression of a pgm2 gene encoding phosphoglucomutase enhanced 2.1 fold increase in crocin-4 from 65.23 ± 2.97μg/L to 136.17 ± 5.89μg/L in flask cultivation. Finally, a batch culture operated under the conditions (20℃, pH 5.5, and a DO of 40%) enhanced up to 204.25 ± 8.83 μg/L of crocin-4 in a bioreactor. To the best of our knowledge, our study presents the first crocin-producing S. cerevisiae cell factory.
Language
eng
URI
https://aurora.ajou.ac.kr/handle/2018.oak/20654
Journal URL
https://dcoll.ajou.ac.kr/dcollection/common/orgView/000000031684
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