Cloning and Expression of Tannase Gene Tan2 from Aspergillus niger

doi:10.13560/j.cnki.biotech.bull.1985.2020-0800

Abstract

Abstract:

Tannase（EC 3.1.1.20）can hydrolyze the ester bond and carboxyphenolic acid bond in tannins to produce gallic acid and corresponding alcohols. It is widely used in food,beverage,feed,pharmacy,medicine,cosmetics and other industries,and also plays an important role in the fermentation of Pu-erh tea. In this work,tannase gene Tan 2 was cloned from Aspergillus niger PU001,which was previously isolated from fermentation in Pu-erh tea. It was ligated with expression vector pCold-I to construct the prokaryotic cold inducible expression system of BL21-pCold I,which was transformed into Escherichia coli BL21. SDS-PAGE and mass spectrometry identification showed that tannase Tan 2 was expressed successfully. This result provides a basis for further survey on the role of tannase in the fermentation of Pu-erh tea and application in other fields.

Key words: tannase, Aspergillus niger, Pu-erh tea

LI Hong-ye, CHEN Li-jiao, LIU Ming-li, GUO Tian-jie, WANG Dao-ping, PAN Ying-hong, ZHAO Ming. Cloning and Expression of Tannase Gene Tan2 from Aspergillus niger[J]. Biotechnology Bulletin, 2021, 37(3): 44-52.

Figures/Tables 10

References 26

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Accession	Blast results（NCBI）
Accession	Description	Organism	Identity
XP_001401809.1	tannase	Aspergillus niger CBS 513.88	100%
EHA20668.1	hypothetical protein	Aspergillus niger ATCC 1015	99.83%
GCB23703.1	tannase	Aspergillus awamori	99.65%
RDK47096.1	tannase and feruloyl esterase	Aspergillus phoenicis ATCC 13157	99.48%
XP_026632695.1	tannase and feruloyl esterase	Aspergillus welwitschiae	99.30%
XP_025515125.1	tannase and feruloyl esterase	Aspergillus piperis CBS 112811	97.91%
XP_025567423.1	tannase and feruloyl esterase	Aspergillus vadensis CBS 113365	97.91%
OJZ87444.1	hypothetical protein	Aspergillus luchuensis CBS 106.47	97.74%
GAA82877.1	tannase and feruloyl esterase	Aspergillus luchuensis IFO 4308	97.56%
OJJ71084.1	hypothetical protein	Aspergillus brasiliensis CBS 101740	97.56%

Accession	Blast results（NCBI）
Accession	Description	Organism	Identity
XP_001401809.1	tannase	Aspergillus niger CBS 513.88	100%
EHA20668.1	hypothetical protein	Aspergillus niger ATCC 1015	99.83%
GCB23703.1	tannase	Aspergillus awamori	99.65%
RDK47096.1	tannase and feruloyl esterase	Aspergillus phoenicis ATCC 13157	99.48%
XP_026632695.1	tannase and feruloyl esterase	Aspergillus welwitschiae	99.30%
XP_025515125.1	tannase and feruloyl esterase	Aspergillus piperis CBS 112811	97.91%
XP_025567423.1	tannase and feruloyl esterase	Aspergillus vadensis CBS 113365	97.91%
OJZ87444.1	hypothetical protein	Aspergillus luchuensis CBS 106.47	97.74%
GAA82877.1	tannase and feruloyl esterase	Aspergillus luchuensis IFO 4308	97.56%
OJJ71084.1	hypothetical protein	Aspergillus brasiliensis CBS 101740	97.56%

	单宁酶/（U·mL^-1）	ΔGA/（mg·mL^-1）	PG/（mg·mL^-1）
样品管	1.02±0.06	0.0034±0.0002	0.0486±0.00115
空白管	—	—	0.0545±0.00309
对照管	—	—	0.0527±0.00274

	单宁酶/（U·mL^-1）	ΔGA/（mg·mL^-1）	PG/（mg·mL^-1）
样品管	1.02±0.06	0.0034±0.0002	0.0486±0.00115
空白管	—	—	0.0545±0.00309
对照管	—	—	0.0527±0.00274