Acetic Acid Addition: Effects on the Production of 2,3-butanediol by Saccharomyces cerevisiae

doi:10.11924/j.issn.1000-6850.casb20190500171

Abstract

Abstract:

To verify the role of acetic acid as a signal molecule, the supernatant of Saccharomyces cerevisiae W141 (the control group) and W141-07 (△aldh6), as well as 1.5 g/L acetic acid were added into S. cerevisiae W141 fermentation broth at the logarithmic phase. The content of 2,3- butanediol(2,3-BD), the activity of acetolactate synthase (ILV2) and 2,3-BD dehydrogenase (BDH1) were detected. The results showed that under the condition of 1.5 g/L acetic acid, the content of 2,3-BD, the ILV2 and BDH1 enzyme activities reached the highest value, which were 3.01±0.04 g/L, 1.41±0.03 U/mg and 0.12±0.002 U/mg, respectively. The acetic acid addition group was significantly higher than those of supernatant addition groups (P<0.01). At the same time, the expressions of ilv2 (24 h) and bdh1 (60 h) genes under the three conditions were determined. After adding W141-07 supernatant, the expression levels of ilv2 and bdh1 genes were down-regulated by 31.6% and 25.0%, respectively. After adding acetic acid with a final concentration 1.5 g/L, the expression levels of ilv2 and bdh1 were up-regulated, which were 4.38 and 1.24 times that of the control, respectively. These results indicate that acetic acid can act as a signal molecule to drive the expression of related genes, thus to improve 2,3-BD production.

Key words: 2,3-butanediol, Saccharomyces cerevisiae, acetic acid, fluorescent quantitative polymerase chain reaction, 2,3-butanediol dehydrogenase, α-acetolactatesynthase

CLC Number:

Q935

Yang Zhiyu, Tong Tianqi, Liu Lei, Ping Wenxiang, Ge Jingping. Acetic Acid Addition: Effects on the Production of 2,3-butanediol by Saccharomyces cerevisiae[J]. Chinese Agricultural Science Bulletin, 2020, 36(21): 104-112.

Figures/Tables 12

References 30

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引物名称	引物序列(5’-3’)
18S-up	TCACC AGGTC CAGAC ACAAT
18S-down	AGCAG ACAAA TCACT CCACC
qilv2-up	CGTCCAATTCCTCTTGCTTC
qilv2-down	ATGGCAATCCCTGTTCTACG
qbdh1-up	TTTGCTGAACAAGTCGTAGTC
qbdh1-down	CCCAGTTTCTTGGCCATTTC

引物名称	引物序列(5’-3’)
18S-up	TCACC AGGTC CAGAC ACAAT
18S-down	AGCAG ACAAA TCACT CCACC
qilv2-up	CGTCCAATTCCTCTTGCTTC
qilv2-down	ATGGCAATCCCTGTTCTACG
qbdh1-up	TTTGCTGAACAAGTCGTAGTC
qbdh1-down	CCCAGTTTCTTGGCCATTTC

时间	组别	CT值		△CT	△△CT	2^-△△^CT
时间	组别	CT-ilv2	CT-18S	△CT	△△CT	2^-△△^CT
24 h	添加S. cerevisiae W141上清液（对照）	23.99±0.32	20.13±0.11	3.77	0	1
	添加S. cerevisiae W141-07上清液	23.76±0.22	19.59±0.16	4.17	0.40	0.76
	添加1.5 g/L乙酸	23.01±0.16	18.97±0.45	4.04	-2.13	4.38

时间	组别	CT值		△CT	△△CT	2^-△△^CT
时间	组别	CT-ilv2	CT-18S	△CT	△△CT	2^-△△^CT
24 h	添加S. cerevisiae W141上清液（对照）	23.99±0.32	20.13±0.11	3.77	0	1
	添加S. cerevisiae W141-07上清液	23.76±0.22	19.59±0.16	4.17	0.40	0.76
	添加1.5 g/L乙酸	23.01±0.16	18.97±0.45	4.04	-2.13	4.38

时间	组别	CT值		△CT	△△CT	2^-△△^CT
时间	组别	CT- bdh1	CT-18S	△CT	△△CT	2^-△△^CT
60 h	添加S. cerevisiae W141上清液（对照）	24.57±0.21	19.03±0.31	5.54	0	1
	添加S. cerevisiae W141-07上清液	24.94±0.32	19.07±0.16	5.87	0.33	0.80
	添加1.5 g/L乙酸	23.82±0.25	19.33±0.41	4.49	-0.31	1.24