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Method and special enzymic preparation for preparing ultra-high maltose syrup

A maltose syrup and ultra-high technology, applied in the field of preparation of ultra-high maltose syrup, can solve problems such as complex process, and achieve the effects of simple process, strong maltose generating capacity and low production cost

Inactive Publication Date: 2011-12-21
江苏锐阳生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] At present, the fungal α-amylase used in the process of preparing high maltose syrup both at home and abroad is Aspergillus oryzae ( A. oryzae ), the disadvantage of using this enzyme is that it must be used in combination with other enzymes, and the prepared high maltose syrup still contains a relatively high concentration of maltotriose (about 10%-20%)
In the production of ultra-high maltose syrup, a variety of enzyme preparations must work together during the saccharification process, and the process is complex

Method used

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  • Method and special enzymic preparation for preparing ultra-high maltose syrup
  • Method and special enzymic preparation for preparing ultra-high maltose syrup
  • Method and special enzymic preparation for preparing ultra-high maltose syrup

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0029] Embodiment 1: the preparation of ultra-high maltose syrup

[0030] Liquefaction: Prepare 100 kg of corn starch into starch milk with a concentration of 10%-30%, adjust the pH to 4.5-6.5, heat to 95-105°C, add high-temperature-resistant acid a-amylase, and the amylase dosage is 2- 8 LU / g dry starch, liquefy for 10-25 minutes, inactivate enzyme at 135°C for 5 minutes, control DE value between 1-8;

[0031] Saccharification: Cool to 55-65°C after liquefaction, add fungal a-amylase RoAmy, the dosage is 4-15 U / g dry starch, during this period, use high-pressure liquid chromatography (HPLC) to measure the formation of glucose, maltose and other oligosaccharides After 30-60 hours of saccharification, decolorization, ion exchange, and vacuum concentration to a concentration of 70%-80%, 125-140 kg of ultra-high maltose syrup with a maltose content of 65%-75% was obtained.

Embodiment 2

[0032] Example 2: Cloning of fungal α-amylase RoAmy

[0033] Extract total RNA from Rhizopus oryzae tissue, using the following oligonucleotides as primers:

[0034] Upstream primer: 5'-ATCATGAAGTCTTTTCTTAAGTCTCCTTTGCA-3',

[0035] Downstream primer: 5'-CTGCCCGGGTTATTTCTTTTGGAA-3'

[0036] Using a reverse transcription PCR kit to amplify to a size of 1.4 kb Ro Amy The cDNA fragment of the gene was recovered and cloned into pMD19, and the recombinant plasmid pMD- Ro Amy , extract the recombinant plasmid and sequence it using T-vector universal primers to determine Ro Amy The cDNA sequence of the gene is SEQ ID NO:1.

Embodiment 3

[0037] Example 3: Construction of Yeast Expression Fungal α-amylase RoAmy Vector

[0038] With the recombinant plasmid pMD- Ro Amy As a template, the following oligonucleotides are used as primers

[0039]Upstream primer: 5'-ATCATGAAGTCTTTTCTTAAGTCTCCTTTGCA-3',

[0040] Downstream primer: 5'-CTGCCCGGGTTATTTCTTTTGGAA-3'

[0041] Using a high-fidelity DNA polymerase ( Pfu DNA polymerase) amplified Ro Amy gene fragments, recovery Ro Amy The gene fragment was inserted into the pPIC9K vector (from Invitrogen) in a blunt-end manner Snap BI site to obtain the yeast expression vector pPIC9K- Ro Amy .

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Abstract

A method and a special enzymic preparation for preparing ultra-high maltose syrup belong to the technical field of starch sugar generation and microbial gene engineering. The method is characterized by using fungusa-amylase RoAmy provided by the invention and using various plant starches as the raw materials and comprises the following steps: liquefying starch milk with high temperature resistantacid a-amylase, and then cooling the liquefied starch milk to 55-65 DEG C, adjusting the pH value to 4.5-6.5, adding the fungusa-amylase RoAmy and preserving heat for 30-60h, thus preparing 65-75% ofultra-high maltose syrup, wherein the gene of the fungusa-amylase RoAmy roots in rhizopus oryzae and the recombinant fungusa-amylase RoAmy is obtained by genetic engineering means. The invention alsoprovides a method for efficiently preparing yeast for the gene of RoAmy. In the method, the ultra-high maltose syrup is obtained by saccharifying starch with the fungusa-amylase RoAmy, and compared with the traditional preparation methods of ultra-high maltose syrup, the method of the invention has the advantages of simple process, low cost, benefit for amplification, etc.

Description

technical field [0001] The present invention relates to a preparation method of ultra-high maltose syrup and its special enzyme preparation, and more specifically relates to a preparation method of ultra-high concentration maltose syrup and the basic characteristics and preparation method of its special enzyme preparation, belonging to the production of starch sugar technology and microbial genetic engineering technology field. Background technique [0002] Maltose syrup is a maltose-based syrup made of starch as raw material, which is hydrolyzed by enzymes or a combination of acid and enzymes. According to the content of maltose, it can be roughly divided into maltose (maltose content 20%-30%), High maltose syrup (maltose content 40%-60%), ultra-high maltose syrup (maltose content greater than 65%) and crystallized maltose, etc. [0003] High maltose syrup has low and mild sweetness, strong palatability, is relatively stable under high temperature and acidic conditions, ha...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N9/30C12N15/56C12N15/63C12P19/14C12R1/685C12R1/645
Inventor 王正祥石贵阳李松
Owner 江苏锐阳生物科技有限公司
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