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Alpha-L-rhamnosidase Rha1 as well as expressed gene and application of alpha-L-rhamnosidase Rha1

A rhamnosidase and gene technology, applied in the fields of genetic engineering and enzyme engineering, can solve problems such as low cost and achieve the effect of reducing energy consumption

Active Publication Date: 2015-01-28
NANJING FORESTRY UNIV
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Problems solved by technology

Aiming at the disadvantages that no α-L-rhamnosidase gene derived from Aspergillus niger has been cloned and expressed, and it is difficult to obtain the pure enzyme of Aspergillus niger α-L-rhamnosidase for the biotransformation of flavonoids at low cost , the present invention provides a kind of α-L-rhamnosidase Rha1 derived from Aspergillus niger and its expression gene and application

Method used

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  • Alpha-L-rhamnosidase Rha1 as well as expressed gene and application of alpha-L-rhamnosidase Rha1
  • Alpha-L-rhamnosidase Rha1 as well as expressed gene and application of alpha-L-rhamnosidase Rha1
  • Alpha-L-rhamnosidase Rha1 as well as expressed gene and application of alpha-L-rhamnosidase Rha1

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Embodiment 1

[0032] 1. Obtaining total RNA from Aspergillus niger

[0033] 1.1 Cultivation of Aspergillus niger

[0034] Aspergillus niger strain (Aspergillus niger) NL-1 was screened and obtained by our laboratory (cloning, expression and directional transformation of Aspergillus niger cbh1 gene and high glucose tolerance bgl1 gene, Li Guoqing, Nanjing Forestry University, master's thesis, 2012), preserved in Nanjing Forestry University For microbial strain collections, the applicant guarantees to release biological materials to the public within twenty years from the date of application. The medium formula of Aspergillus niger strain (Aspergillus niger) is: glucose 30g / L, K 2 HPO 4 ·3H 2 O1g / L, KCl0.5g / L, MgSO 4 0.5g / L, FeSO 4 0.01g / L, NaNO 30.2g / L, adjust the pH to 4.8, inoculate fresh Aspergillus niger spore suspension, culture at 180rpm at 30°C for 3-4 days and collect mycelium by filtration.

[0035] 1.2 Extraction of total RNA from Aspergillus niger

[0036] Take the collect...

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Abstract

The invention relates to alpha-L-rhamnosidase Rha1 as well as an expressed gene and an application of the alpha-L-rhamnosidase Rha1. An amino acid sequence is shown in SEQIDNO.2, a nucleotide sequence of the gene coding alpha-L-rhamnosidase Rha1 is shown in SEQIDNO.1. 1.5g / L of rutin is hydrolyzed for 20min by the alpha-L-rhamnosidase Rha1 under the conditions that the dosage of enzyme is 13U / ml, pH value is 6.5 and the temperature is 35 DEG C, and the hydrolyzing efficiency is more than 98%. Flavonoids compounds, such as rutin, aringin and the like can biologically catalyzed and converted by alpha-L-rhamnosidase Rha1 at a low temperature, and the energy consumption is greatly reduced.

Description

technical field [0001] The invention belongs to the technical field of genetic engineering and enzyme engineering, and specifically relates to α-L-rhamnosidase Rha1 derived from Aspergillus niger and its expression gene and application. Background technique [0002] α-L-rhamnosidase (α-L-rhamnosidase, EC3.2.1.40) widely exists in animals, plants and microorganisms, and can specifically hydrolyze many glycosides, such as naringin, rutin, orange The terminal α-L-rhamnosyl of glucosides such as dermoside can be used in food, medicine and other fields. The main functions and functions include: [0003] (1) Debittering effect: flavanone glycosides are the main bitter substances in citrus. α-L-rhamnosidase can act on flavanone glycosides, and after removing a rhamnose molecule, transform It is a monoglycoside, and its bitterness is one-third of the original. [0004] (2) Flavoring effect: There are a large number of bonded aromatic substances in glucose. The bonded aromatic sub...

Claims

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Application Information

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IPC IPC(8): C12N9/24C12N15/56C12N15/63C12N15/66C12N1/21C12P19/60
CPCC12N9/2402C12P17/06C12Y302/0104
Inventor 赵林果裴建军冯翊阳汤锋岳永德操海群余晓丹
Owner NANJING FORESTRY UNIV
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