CN101948888B - Method of utilizing microbiological debranching for preparing banana resistant starch - Google Patents
Method of utilizing microbiological debranching for preparing banana resistant starch Download PDFInfo
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- CN101948888B CN101948888B CN 201010267475 CN201010267475A CN101948888B CN 101948888 B CN101948888 B CN 101948888B CN 201010267475 CN201010267475 CN 201010267475 CN 201010267475 A CN201010267475 A CN 201010267475A CN 101948888 B CN101948888 B CN 101948888B
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Abstract
The invention discloses a method of utilizing microbiological debranching for preparing banana resistant starch. The preparation method comprises the following processes: pretreating raw material, preparing a culture medium, performing slant culture, preparing spore suspension and bacterial suspension, performing fermentation culture and metabolism debranching, breaking cell walls, separating resistant starch and performing microwave drying. The method of the invention uses the selected Bacillus subtilis and aspergillus niger to metabolize alpha-amylase, beta-amylase and debranching enzyme and perform the enzymolysis of macromolecular amylose, thus obtaining micromolecular amylose and debranching amylose; and the method of the invention improves the commonly used preparation method of the resistant starch, wherein cell walls are broken in the fermentation culture period to release more in vivo enzyme, thus the yield of the resistant starch is significantly increased and accounts for more than 40% of the resistant starch amount of the original banana fruit, and the purity of the resistant starch is up to 99.8%.
Description
Technical field
The present invention relates to a kind of preparation method of banana Resistant starch, particularly utilize the method for microbiological debranching for preparing banana resistant starch, belong to biological technical field.
Background technology
Starch is divided into amylose starch and amylopectin from structure, the polymerization degree of amylose starch is generally at 500～6000 glucosyl residues, amylopectin is made of A chain, B chain and C chain, and the C chain is main chain, and a reduction end is arranged, the polymerization degree is generally at 60 more than glucosyl residue, the B chain is the side chain on the C chain, and the polymerization degree is generally 45～55 glucosyl residues, and the A chain is the side chain on the B chain, the polymerization degree is generally 14～18 glucosyl residues, and amylopectin is the amylose starch of short chain after taking off and propping up.Starch is from being divided in nature three types: the one, and quick amyloclastic (rapid digested starch); The 2nd, amyloclastic (slowly digested starch) at a slow speed; The 3rd, Resistant starch (resistant starch) is called for short RS, and Resistant starch is defined as " starch that is not absorbed by the healthy individuals small intestine and the general name of degraded product thereof ".
Resistant starch has different physiological roles, and it can promote the growth and breeding of intestinal beneficial bacterium clump, is a kind of bifidus bacillus multiplicaiton factor, can increase the ight soil capacity, plays an important role to preventing constipation, typhlitis and hemorrhoid.Resistant starch ferments in large intestine or the part fermentation, produces the volatility short chain fatty acid, as butyric acid, and propionic acid, acetic acid etc. reduce the pH value in large intestine, reduce the colorectal carcinoma sickness rate, suppress the growth and breeding of pathogenic bacterium.Resistant starch is easier of the microbial fermentation in large intestine than food fibre, compare with other food fibre, the butyric acid percentage of Resistant starch fermentation institute output is the highest, takes in Resistant starch food, can reduce the rising of blood sugar after meal and the secretion of promotion Regular Insulin, improve simultaneously lipid and consist of.Resistant starch is comprised of crystallizing field and pars amorpha, and crystallizing field mainly is made of the amylose starch of crystallization.The ability of Resistant starch antiamylase class digestion comes from the amylose starch crystal.Amylose starch amylose molecule under certain condition is close to each other, intermolecularly forms duplex by hydrogen bond, and duplex mutually superposes and forms the amylose starch crystal, makes long chain amylose become short chain amylose, and short chain amylose easily forms Resistant starch.
" fine jade state institute journal " the 16th the 2nd phase of volume 55-60 page in 2009 has been delivered " blue or green banana Resistant starch optimization for extracting condition ", its method is by factors such as enzyme addition, hydrolysis time, temperature, pectinase activity to be affected, the suitableeest hydrolysising condition of determining the blue or green banana starch of extraction is solid-liquid ratio 1: 2, temperature 45 C, enzymolysis time 50min, but its starch pick-up rate is only 4.67%; Patent Office of State Intellectual Property Office disclosed " preparation method and the application of natural banana Resistant starch RS2 " patent application (notification number: CN101792783A), its method is banana blanching, peeling, allotment, access aspergillus niger and Pseudomonas aeruginosa fermentation preparation banana Resistant starch, but do not carry out cell wall breaking, above-mentioned two kinds of methods are all the banana peelings, cause the waste and secondary pollution, and the pick-up rate of Resistant starch and purity low.And banana is carried out full fruit making beating, by aspergillus niger and subtilis metabolism debranching factor, the method that the fermentation culture later stage uses penicillin to carry out cell wall breaking has not yet to see report.
Summary of the invention
The objective of the invention is to prepare the banana Resistant starch by microbial fermentation, subtilis and aspergillus niger difference metabolism α-amylase, beta-amylase and debranching factor by seed selection, be macromole amylose starch enzymolysis the small molecules amylose starch, amylopectin is sloughed side chain.
The present invention prepares to complete by the following method:
A. raw materials pretreatment
Immature blue or green banana is cleaned up, soaked 10-20 minute in the citric acid colour protecting liquid, then full fruit making beating, cross the 50-120 eye mesh screen and separate robust fibre, obtains banana pulp;
B. prepare substratum
The weight part proportioning of each component of substratum is:
Banana pulp 10-30, (NH
4)
2SO
40.5-1, NaH
2PO
41-2, MgSO
47H
2O 0.1-1,
Distilled water 80-90,
With banana pulp, (NH
4)
2SO
4, NaH
2PO
4, MgSO
47H
2O mixes, then adds distilled water to carry out constant volume, and it is stand-by that the triangular flask of packing into, microwave disinfection 10-30 form substratum second;
C. slant culture and spore suspension and bacterium liquid suspension preparation
Get aspergillus niger access PDA slant medium, subtilis access extractum carnis slant medium, inoculation is placed in incubator 30 ℃ and cultivated 72 hours respectively, then with the spore on sterilized water washing PDA slant medium and the bacterium colony on the extractum carnis slant medium, make respectively spore suspension and bacterium liquid suspension;
D. fermentation culture and metabolism are taken off and are propped up
The weight part proportioning of each component of fermentation culture is:
Substratum 90-120, spore suspension 0.15-0.5, bacterium liquid suspension 0.15-0.5,
The penicillin 0.000020-0.000040 of 25u/mL,
Add respectively spore suspension and bacterium liquid suspension in substratum, then be placed on shaking table, the control rotating speed is 90-180r/min, 30 ℃-50 ℃ fermentation culture 10-30 hour;
E. cell wall breaking
When fermentation culture proceeds to 16-36 hour, then add penicillin in substratum, make cell wall breaking, discharge more body endoenzyme, then continued fermentation culture 1-6 hour;
F. separation resistance starch
Adopt high speed three-phase decanter separating machine to isolate wet Resistant starch, separator drum speed 10000-14000r/min, differential 30-50r/min;
G. microwave drying
100-120 ℃ of microwave drying temperature, gets Resistant starch at 50-120 second time of drying.
The present invention has improved the preparation method of Resistant starch commonly used at present, carry out cell wall breaking during fermentation culture, discharge more body endoenzyme, the pick-up rate of Resistant starch is significantly improved, reach more than 40% of former banana fruit, the quality purity of Resistant starch reaches 99.8% simultaneously.
Embodiment
By following examples, the present invention is described in further detail:
Embodiment one:
A. raw materials pretreatment
After selecting immature blue or green banana to clean, soaked 10 minutes in the colour protecting liquid citric acid, full fruit making beating is crossed 100 eye mesh screens and is isolated robust fibre, obtains banana pulp;
B. prepare substratum
Get banana pulp weight part 30, (NH
4)
2SO
4Weight part 0.5, NaH
2PO
4Weight part 1, MgSO
47H
2O weight part 0.1 mixes, and adds distilled water weight part 80 to carry out constant volume, the triangular flask of packing into, and it is stand-by that microwave disinfection formed substratum in 30 seconds;
C. slant culture and spore suspension and bacterium liquid suspension preparation
Get aspergillus niger access PDA slant medium, subtilis access extractum carnis slant medium, inoculation is placed in incubator 30 ℃ and cultivated 72 hours respectively, then with the spore on sterilized water washing PDA slant medium and the bacterium colony on the extractum carnis slant medium, make respectively spore suspension and bacterium liquid suspension;
D. fermentation culture
Each 0.15 adds in above-mentioned substratum to get respectively spore suspension and bacterium liquid suspension weight part, then is placed on shaking table, and the control rotating speed is 180r/min, 30 ℃ of fermentation culture 10 hours;
E. cell wall breaking
Fermentation culture proceeds to 10 hours, adds the penicillin weight part 0.000025 of 25u/mL in the substratum, makes cell wall breaking, discharges more body endoenzyme, then continues fermentation culture 4 hours;
F. separation resistance starch
Adopt high speed three-phase decanter separating machine to isolate wet Resistant starch, separator drum speed 14000r/min, differential 35r/min;
G. microwave drying
120 ℃ of microwave drying temperature, 50 seconds time of drying, the Resistant starch pick-up rate is 43% of former banana fruit.
Embodiment two:
A. raw materials pretreatment
After selecting immature blue or green banana to clean, soaked 10 minutes in the colour protecting liquid citric acid, full fruit making beating is crossed 80 eye mesh screens and is isolated robust fibre, obtains banana pulp;
B. prepare substratum
Get banana pulp weight part 20, (NH
4)
2SO
4Weight part 0.5, NaH
2PO
4Weight part 1, MgSO
47H
2O weight part 0.1 adds distilled water weight part 86 to carry out constant volume, the triangular flask of packing into, and it is stand-by that microwave disinfection formed substratum in 30 seconds;
C. slant culture and spore suspension and bacterium liquid suspension preparation
Get aspergillus niger access PDA slant medium, subtilis access extractum carnis slant medium, inoculation is placed in incubator 30 ℃ and cultivated 72 hours respectively, then with the spore on sterilized water washing PDA slant medium and the bacterium colony on the extractum carnis slant medium, make respectively spore suspension and bacterium liquid suspension;
D. fermentation culture
Each 0.2 adds in above-mentioned substratum to get respectively spore suspension and bacterium liquid suspension weight part, then is placed on shaking table, and the control rotating speed is 180r/min, 40 ℃ of fermentation culture 20 hours;
E. cell wall breaking
Fermentation culture proceeds to 20 hours, adds the penicillin weight part 0.000020 of 25u/mL in the substratum, makes cell wall breaking, discharges more body endoenzyme, then continues fermentation culture 4 hours;
F. adopt high speed three-phase decanter separating machine to isolate wet Resistant starch, separator drum speed 13000r/min, differential 30r/min;
G. microwave drying
160 ℃ of microwave drying temperature, 60 seconds time of drying, the Resistant starch pick-up rate is 46% of former banana fruit.
Embodiment three:
A. raw materials pretreatment
After selecting immature blue or green banana to clean, soaked 10 minutes in the colour protecting liquid citric acid, full fruit making beating is crossed 110 eye mesh screens and is isolated robust fibre, obtains banana pulp;
B. prepare substratum
Get banana pulp weight part 20, (NH
4)
2SO
4Weight part 0.5, NaH
2PO
4Weight part 1, MgSO
47H
2O weight part 0.1 adds distilled water weight part 85 to carry out constant volume, the triangular flask of packing into, and it is stand-by that microwave disinfection formed substratum in 30 seconds;
C. slant culture and spore suspension and bacterium liquid suspension preparation
Get aspergillus niger access PDA slant medium, subtilis access extractum carnis slant medium, inoculation is placed in incubator 30 ℃ and cultivated 72 hours respectively, then with the spore on sterilized water washing PDA slant medium and the bacterium colony on the extractum carnis slant medium, make respectively spore suspension and bacterium liquid suspension;
D. fermentation culture
Each 0.3 adds in above-mentioned substratum to get respectively spore suspension and bacterium liquid suspension weight part, then is placed on shaking table, and the control rotating speed is 180r/min, 38 ℃ of fermentation culture 30 hours;
E. cell wall breaking
Fermentation culture proceeds to 30 hours, adds the penicillin weight part 0.000040 of 25u/mL in substratum, then continues fermentation culture 6 hours;
F. adopt high speed three-phase decanter separating machine to isolate wet Resistant starch, separator drum speed 10000r/min, differential 20r/min;
G. microwave drying
100 ℃ of microwave drying temperature, 90 seconds time of drying, the Resistant starch pick-up rate is 44% of former banana fruit.
Embodiment four:
A. raw materials pretreatment
After selecting immature blue or green banana to clean, soaked 10 minutes in the colour protecting liquid citric acid, full fruit making beating is crossed 600 eye mesh screens and is isolated robust fibre, obtains banana pulp;
B. prepare substratum
Get banana pulp weight part 20, (NH
4)
2SO
4Weight part 0.5, NaH
2PO
4Weight part 1, MgSO
47H
2O weight part 0.1 adds distilled water weight part 90 to carry out constant volume, the triangular flask of packing into, and it is stand-by that microwave disinfection formed substratum in 20 seconds;
C. slant culture and spore suspension and bacterium liquid suspension preparation
Get aspergillus niger access PDA slant medium, subtilis access extractum carnis slant medium, inoculation is placed in incubator 30 ℃ and cultivated 72 hours respectively, then with the spore on sterilized water washing PDA slant medium and the bacterium colony on the extractum carnis slant medium, make respectively spore and suspension bacterium liquid suspension;
D. fermentation culture
Each 0.45 adds/enters in above-mentioned substratum to get respectively spore suspension and bacterium liquid suspension weight part, then is placed on shaking table, and the control rotating speed is 180r/min, 48 ℃ of fermentation culture 35 hours;
E. cell wall breaking
Fermentation culture proceeds to 35 hours, adds the penicillin weight part 0.000030 of 25u/mL in substratum, then continues fermentation culture 5 hours;
F. adopt high speed three-phase decanter separating machine to isolate wet Resistant starch, separator drum speed 12000r/min, differential 25r/min;
G. microwave drying
80 ℃ of microwave drying temperature, 100 seconds time of drying, the Resistant starch pick-up rate is 42% of former banana fruit.
Claims (2)
1. utilize the method for microbiological debranching for preparing banana resistant starch, it is characterized in that completing by following concrete grammar preparation:
A. after selecting immature blue or green banana to clean, soaked 10 minutes in the colour protecting liquid citric acid, full fruit making beating is crossed 100 eye mesh screens and is isolated robust fibre, obtains banana pulp;
B. get banana pulp weight part 30, (NH
4)
2SO
4Weight part 0.5, NaH
2PO
4Weight part 1, MgSO
47H
2O weight part 0.1 mixes, and adds distilled water weight part 80 to carry out constant volume, the triangular flask of packing into, and it is stand-by that microwave disinfection formed substratum in 30 seconds;
C. get aspergillus niger access PDA slant medium, subtilis access extractum carnis slant medium, inoculation is placed in incubator 30 ℃ and cultivated 72 hours respectively, then with the spore on sterilized water washing PDA slant medium and the bacterium colony on the extractum carnis slant medium, make respectively spore suspension and bacterium liquid suspension;
D. each 0.15 adds in above-mentioned substratum to get respectively spore suspension and bacterium liquid suspension weight part, then is placed on shaking table, and the control rotating speed is 180r/min, 30 ℃ of fermentation culture 10 hours;
E. fermentation culture proceeds to 10 hours, adds the penicillin weight part 0.000025 of 25u/mL in the substratum, makes cell wall breaking, discharges more body endoenzyme, then continues fermentation culture 4 hours;
F. adopt high speed three-phase decanter separating machine to isolate wet Resistant starch, separator drum speed 14000r/min, differential 35r/min;
G. the microwave drying temperature is 120 ℃, and 50 seconds time of drying, the Resistant starch pick-up rate is 43% of former banana fruit.
2. utilize the method for microbiological debranching for preparing banana resistant starch, it is characterized in that completing by following concrete grammar preparation:
A. after selecting immature blue or green banana to clean, soaked 10 minutes in the colour protecting liquid citric acid, full fruit making beating is crossed 80 eye mesh screens and is isolated robust fibre, obtains banana pulp;
B. get banana pulp weight part 20, (NH
4)
2SO
4Weight part 0.5, NaH
2PO
4Weight part 1, MgSO
47H
2O weight part 0.1 adds distilled water weight part 86 to carry out constant volume, the triangular flask of packing into, and it is stand-by that microwave disinfection formed substratum in 30 seconds;
C. get aspergillus niger access PDA slant medium, subtilis access extractum carnis slant medium, inoculation is placed in incubator 30 ℃ and cultivated 72 hours respectively, then with the spore on sterilized water washing PDA slant medium and the bacterium colony on the extractum carnis slant medium, make respectively spore suspension and bacterium liquid suspension;
D. each 0.2 adds in above-mentioned substratum to get respectively spore suspension and bacterium liquid suspension weight part, then is placed on shaking table, and the control rotating speed is 180r/min, 40 ℃ of fermentation culture 20 hours;
E. fermentation culture proceeds to 20 hours, adds the penicillin weight part 0.000020 of 25u/mL in the substratum, makes cell wall breaking, discharges more body endoenzyme, then continues fermentation culture 4 hours;
F. adopt high speed three-phase decanter separating machine to isolate wet Resistant starch, separator drum speed 13000r/min, differential 30r/min;
G. the microwave drying temperature is 160 ℃, and 60 seconds time of drying, the Resistant starch pick-up rate is 46% of former banana fruit.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 201010267475 CN101948888B (en) | 2010-08-30 | 2010-08-30 | Method of utilizing microbiological debranching for preparing banana resistant starch |
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 201010267475 CN101948888B (en) | 2010-08-30 | 2010-08-30 | Method of utilizing microbiological debranching for preparing banana resistant starch |
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| CN101948888B true CN101948888B (en) | 2013-05-15 |
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| DE102013112165A1 (en) * | 2013-11-05 | 2015-05-07 | Gea Mechanical Equipment Gmbh | Process for obtaining starch |
| CN105852138B (en) * | 2016-03-31 | 2018-11-20 | 天蕉健康食品（广东）有限公司 | A kind of extracting method of banana natural resistance starch |
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| CN101792783B (en) * | 2010-02-25 | 2012-08-22 | 华南农业大学 | Preparation method and application of banana natural resistant starch RS2 |
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