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Maltotriose preparation method with starch as raw material and special fungal alpha-amylase thereof

A maltotriose and amylase technology is applied in the field of the preparation of maltotriose, the preparation method of maltotriose and its special fungal alpha-amylase, and achieves the effects of low production cost and simple process

Inactive Publication Date: 2013-04-03
绿爱控股股份有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Later, the α-amylase gene ( SF ) has been studied in hosts such as bacteria and Saccharomyces cerevisiae, but no recombinant Pichia pastoris expression SF A report on the phenomenon of massive accumulation of maltotriose in the products obtained by hydrolyzing starch with genes and recombinant enzymes

Method used

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  • Maltotriose preparation method with starch as raw material and special fungal alpha-amylase thereof
  • Maltotriose preparation method with starch as raw material and special fungal alpha-amylase thereof
  • Maltotriose preparation method with starch as raw material and special fungal alpha-amylase thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0068] Weigh 10 g of cornstarch, use tap water to prepare starch milk with a concentration not higher than 5%, heat it to 80°C, and keep it warm for 10 minutes. The above starch mash was cooled to 45 °C, and 5% hydrochloric acid solution was added to adjust the pH to 4.5, and SfA was added in an amount of 0.15 U / g dry starch, and kept for 6 hours. like Figure 1~3 Shown is the analysis result of sugar in the obtained starch hydrolyzate, wherein the obtained maltotriose accounts for about 52% of the total sugar concentration, and maltose accounts for about 39%. Similar results were obtained using potato starch, sweet potato starch, glutinous rice starch and wheat starch as raw materials, and the content of maltotriose in the obtained hydrolyzate was between 46% and 55% of the total sugar.

[0069] Embodiment 2: Extraction of maltotriose

Embodiment 2

[0070] (1) Fill the chromatographic glass column with activated carbon, and add ordinary tap water from the top to submerge the activated carbon. Add the syrup obtained in Example 1 from the top of the chromatographic column, collect the liquid flowing out from the bottom of the chromatographic column, and detect the reducing sugar content in the effluent liquid. When the reducing sugar content reaches 0.2% in the liquid, it shows that the adsorption capacity of activated carbon has reached saturation. Stop Add sugar solution. Vent the liquid in the activated carbon column. The effluent solution is mainly glucose.

[0071] (2) Close the outlet below the activated carbon column, add 4.5%-5.0% alcohol solution from the upper part of the activated carbon column, until the alcohol solution submerges the activated carbon, and equilibrate for 30 min. Open the outlet below the activated carbon column to release the alcohol solution, which is mainly glucose and maltose.

[0072] (3...

Embodiment 3

[0075] According to the method introduced in the literature [Zhou Xiaoling, Shen Wei, Rao Zhiming, Wang Zhengxiang, Zhuge Jian. A method for rapid extraction of fungal chromosomal DNA. Microbiology Bulletin, 2004, 31(4): 89-92] to extract button capsule and compound membrane Saccharomyces ( Saccharomycopsis fibuligera ) CICIM Y1037 chromosomal DNA.

[0076] According to the α-amylase gene sequence [Itoh T, Yamashita I and Fukui S. Nucleotide sequence of the alpha-amylase gene (ALP1) in the yeast published by Itoh et al. Saccharomycopsis fibuligera . FEBS Lett, 1987, 219(2): 339-342.] Designing primers:

[0077] 5'-AATTACCGTC TAGACAACCA GTGACTCTAT TC-3' (SEQ ID NO: 2),

[0078] 5'-AATTACCGTC TAGATTATGA ACAAATGTCA GAAG-3' (SEQ ID NO: 3)

[0079] Using the above-mentioned extracted chromosomal DNA of S. forticulata as a template, using the above-mentioned primers, PCR amplification was performed with Ex Taq enzyme to obtain the mature peptide coding region of the α-amylase ge...

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Abstract

The invention discloses a maltotriose preparation method with starch as a raw material and a special fungal alpha-amylase thereof and belongs to the technical field of starch sugar manufacturing and enzyme engineering. The preparation method disclosed by the invention comprises the following steps: based on a variety of plant starches as raw materials, carrying out liquefaction and saccharification by utilizing fungal alpha-amylase SfA to obtain the syrup in which maltotriose accounts for 46% to 55% of the total sugar; and carrying out activated carbon adsorption and alcohol elution to obtain the syrup in which maltotriose accounts for not less than 90% of the total sugar. The invention further provides an efficient preparation method of fungal alpha-amylase SfA. The fungal alpha-amylase SfA gene is derived from sacchromycopsis fibuligera CICIMY2037, recombinant Pichia pastoris is obtained by a genetic recombination method, and the SfA efficient preparation is carried out through the recombinant strain. In comparison with the prior method, the preparation method of maltotriose with fungal alpha-amylase SfA has the advantages of low cost of raw materials, independence on starch liquefied enzyme and starch debranching enzyme, simple process and easiness in enlargement.

Description

technical field [0001] The invention discloses a maltotriose preparation method using starch as a raw material and a special fungal α-amylase thereof, relates to a maltotriose preparation method, and belongs to the technical field of starch sugar production and enzyme engineering. Background technique [0002] Maltotriose is an oligosaccharide composed of glucose units connected mainly through α-1,4-glycosidic bonds. Adding maltotriose to foods such as cakes can prevent starch from aging and maintain moisture. The preparation method of maltotriose is mainly divided into two types. One is to use polymaltotriose, namely pullulan, as raw material, which is obtained by decomposition with pullulan enzyme (ZL200910015419.3). Due to the current production of pullulan The cost is higher, so the cost of preparing maltotriose is also higher; the other is obtained by decomposing starch as raw material through maltotriose amylase, because maltotriose amylase is an exo-amylase , so in p...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P19/14C12N1/19C12N15/81C12N9/30C12R1/84C12R1/645
Inventor 沈微郭玉婉黄雯雯樊游陈献忠
Owner 绿爱控股股份有限公司
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