CN101979642A - Method for use in ultrasonic wave-assisted cellulose enzymatic hydrolysis process - Google Patents
Method for use in ultrasonic wave-assisted cellulose enzymatic hydrolysis process Download PDFInfo
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- CN101979642A CN101979642A CN 201010520427 CN201010520427A CN101979642A CN 101979642 A CN101979642 A CN 101979642A CN 201010520427 CN201010520427 CN 201010520427 CN 201010520427 A CN201010520427 A CN 201010520427A CN 101979642 A CN101979642 A CN 101979642A
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Abstract
The invention discloses a method for use in an ultrasonic wave-assisted cellulose enzymatic hydrolysis process, which comprises the following steps: naturally drying wheat straw in the air and crushing the wheat straw into 40-mesh powder for later use; inserting an ultrasonic reactor probe to a fixed depth in reaction solution to perform ultrasonic wave-assisted enzymatic hydrolysis under the conditions below: the substrate concentration is 50g/L, the composite plant hydrolytic enzyme concentration is 0.5fbg/g, the enzymatic hydrolysis temperature is 35 to 60 DEG C, the pH value is 4.4 to 5.2 and the ultrasonic frequency is 24kHz; applying ultrasonic waves for 0 to 40 minutes under a condition that the ultrasonic power is 0 to 60W, stopping the application of the ultrasonic waves and continuing the enzymatic hydrolysis for 4 hours; and centrifuging the reaction solution after enzymatic hydrolysis and obtaining supernate. In the invention, zymologic bioengineering technology and ultrasonic wave technology are combined, so the conversion rate of reduced sugar after the enzymatic hydrolysis of the cellulose in the straw is improved greatly. The ultrasonic wave-assisted cellulose enzymatic hydrolysis serving as an effective method for improving the conversion rate of the reduced sugar after enzymatic hydrolysis has the advantages of simplicity, safety, convenience, quickness and the like.
Description
Technical field
The present invention relates to the influence of the auxiliary stalk cellulose hydrolytic process of ultrasonic wave, belong to technical field of bioengineering.
Background technology
Stalk fermentation is produced alcohol, at first stalk cellulose enzymolysis under the effect of cellulase will be become fermentable sugar, in the zymic effect sugar is converted into alcohol then.In process of production, improving utilization ratio and the hydrolysis efficiency and the transformation efficiency of cellulase, thereby reduce the cost of stalk fermentation alcohol, is the key issue that stalk fermentation fuel alcohol technology realizes industrialization.
In enzymolysis, zymin is the prerequisite that enzyme digestion reaction carries out with fully contacting of substrate, but because the compactness of cellulosic structure, enzymolysis process is subjected to its structural mechanical obstacles, thereby needs auxiliary other means to promote the media reaction to carry out.Supplementary means commonly used at present has: product separation, tensio-active agent etc.Because hyperacoustic characteristic has also caused extensive studies interest.
At present, ultrasonic wave promotes that enzymolysis has been a kind of emerging method, by hyperacoustic reinforcing mass transfer, cavatition etc., promotes the diffusion of enzyme molecule in system, thus the contact of enzyme-to-substrate, the carrying out of promotion enzymolysis.
Summary of the invention
At the lower deficiency of plant fiber material utilization ratio, the purpose of this invention is to provide a kind of technology cellulase hydrolysis efficient is strengthened, for the development that promotes the cellulase hydrolysis technology provides theoretical foundation.
The object of the present invention is achieved like this: the method for the plain enzymolysis process of ultrasonic wave auxiliary fiber comprises the steps:
(1) feedstock production: wheat stalk is behind natural air drying, and it is standby to be crushed to 40 orders;
(2) the auxiliary stalk cellulose hydrolysis of ultrasonic wave: under concentration of substrate 50g/L, composite plant lytic enzyme concentration 0.5fbg/g, hydrolysis temperature 35-60 ℃, the condition of pH value 4.4-5.2, ultrasonic frequency 24kHz, ultrasonic reactor popped one's head in carry out ultrasonic assistance enzymolysis in the insertion reaction liquid, the depth of penetration of wherein popping one's head in immobilizes; At ultrasonic power is to stop ultrasonicly under the 0-60W behind the ultrasonic 0-40min, continues enzymolysis, and enzymolysis time is 4h;
(3) behind the enzymolysis with the reaction solution centrifugation, supernatant liquor.
The present invention combines the zymetology biotechnology with ultrasonic technology, conversion of reduced sugar rate behind the bigger raising stalk cellulose enzymolysis.With respect to the conversion rate of reducing sugar that does not add the auxiliary stalk cellulose enzymolysis of ultrasonic wave, maximum can improve 114%.
The auxiliary stalk cellulose enzymolysis of ultrasonic wave as a kind of effective raising enzymolysis after the conversion rate of reducing sugar method,, safety simple, wait advantage easily and fast because of it, will in the development of industrial sectorization, be widely used.
Embodiment
The method of the conversion rate of reducing sugar among the present invention behind the mensuration enzymolysis is as follows: with the reaction solution centrifugation, supernatant liquor is surveyed its total reducing sugars content with the DNS method behind the end enzymolysis.Conversion rate of reducing sugar is calculated as follows:
Mierocrystalline cellulose and hemicellulose total amount in transformation efficiency (%)=(reducing sugar total amount * 0.9 * 100)/substrate.
Composite plant lytic enzyme Viscozyme L(enzyme used in the present invention 100fbg/g alive) be to originate from Novozymes Company, purchase in Wuxi City and unite permanent continent chemical industry company limited, stalk is taken from Zhengjiang City, Jiangsu Dantu District, the ultrasonic promotion auxanograph of CY-5D type is purchased the biotech inc in Ning Xinzhi.
Comparative Examples:
(1) feedstock production: wheat stalk is behind natural air drying, and it is standby to be crushed to 40 orders.
(2) stalk cellulose hydrolysis: carry out the stalk cellulose enzymolysis under the condition of concentration of substrate 50g/L, 50 ℃ of temperature, pH value 4.8, enzymolysis time is 4h.
(3) measure conversion rate of reducing sugar behind the enzymolysis at last: finish behind the enzymolysis the reaction solution centrifugation, supernatant liquor is surveyed its total reducing sugars content with the DNS method, and calculating conversion rate of reducing sugar by formula is 6.81%.
Embodiment one:
(1) feedstock production: wheat stalk is behind natural air drying, and it is standby to be crushed to 40 orders.
(2) the auxiliary stalk cellulose hydrolysis of ultrasonic wave: under the condition of concentration of substrate 50g/L, 50 ℃ of hydrolysis temperatures, pH value 4.8, ultrasonic frequency 24kHz, ultrasonic reactor popped one's head in carry out ultrasonic enzymolysis (depth of penetration of popping one's head in immobilizes) in the insertion reaction liquid.At ultrasonic power is to stop ultrasonicly under the 30W behind the ultrasonic 5min, continues enzymolysis, and enzymolysis time is 4h.
(3) measure conversion rate of reducing sugar behind the enzymolysis at last: finish behind the enzymolysis the reaction solution centrifugation, supernatant liquor is surveyed its total reducing sugars content with the DNS method.Calculating conversion rate of reducing sugar by formula is 11.24%, than the raising 65% that does not add the auxiliary stalk cellulose hydrolysis of ultrasonic wave.
Embodiment two:
(1) feedstock production: wheat stalk is behind natural air drying, and it is standby to be crushed to 40 orders.
(2) the auxiliary stalk cellulose hydrolysis of ultrasonic wave: under the condition of concentration of substrate 50g/L, 50 ℃ of hydrolysis temperatures, pH value 4.8, ultrasonic frequency 24kHz, ultrasonic reactor popped one's head in carry out ultrasonic enzymolysis (depth of penetration of popping one's head in immobilizes) in the insertion reaction liquid.At ultrasonic power is to stop ultrasonicly under the 30W behind the ultrasonic 40min, continues enzymolysis, and enzymolysis time is 4h.
(3) measure conversion rate of reducing sugar behind the enzymolysis at last: finish behind the enzymolysis the reaction solution centrifugation, supernatant liquor is surveyed its total reducing sugars content with the DNS method.Calculating conversion rate of reducing sugar by formula is 12.19%, than the raising 79% that does not add the auxiliary stalk cellulose hydrolysis of ultrasonic wave.
Embodiment three:
(1) feedstock production: wheat stalk is behind natural air drying, and it is standby to be crushed to 40 orders.
(2) the auxiliary stalk cellulose hydrolysis of ultrasonic wave: under the condition of concentration of substrate 50g/L, 35 ℃ of hydrolysis temperatures, pH value 4.8, ultrasonic frequency 24kHz, ultrasonic reactor popped one's head in carry out ultrasonic enzymolysis (depth of penetration of popping one's head in immobilizes) in the insertion reaction liquid.At ultrasonic power is to stop ultrasonicly under the 30W behind the ultrasonic 10min, continues enzymolysis, and enzymolysis time is 4h.
(3) measure conversion rate of reducing sugar behind the enzymolysis at last: finish behind the enzymolysis the reaction solution centrifugation, supernatant liquor is surveyed its total reducing sugars content with the DNS method.Calculating conversion rate of reducing sugar by formula is 6.92%, than the raising 1.62% that does not add the auxiliary stalk cellulose hydrolysis of ultrasonic wave.
Embodiment four:
(1) feedstock production: wheat stalk is behind natural air drying, and it is standby to be crushed to 40 orders.
(2) the auxiliary stalk cellulose hydrolysis of ultrasonic wave: under the condition of concentration of substrate 50g/L, 60 ℃ of hydrolysis temperatures, pH value 4.8, ultrasonic frequency 24kHz, ultrasonic reactor popped one's head in carry out ultrasonic enzymolysis (depth of penetration of popping one's head in immobilizes) in the insertion reaction liquid.At ultrasonic power is to stop ultrasonicly under the 30W behind the ultrasonic 10min, continues enzymolysis, and enzymolysis time is 4h.
(3) measure conversion rate of reducing sugar behind the enzymolysis at last: finish behind the enzymolysis the reaction solution centrifugation, supernatant liquor is surveyed its total reducing sugars content with the DNS method.Calculating conversion rate of reducing sugar by formula is 9.65%, than the raising 41.7% that does not add the auxiliary stalk cellulose hydrolysis of ultrasonic wave.
Embodiment five:
(1) feedstock production: wheat stalk is behind natural air drying, and it is standby to be crushed to 40 orders.
(2) the auxiliary stalk cellulose hydrolysis of ultrasonic wave: under the condition of concentration of substrate 50g/L, 50 ℃ of hydrolysis temperatures, pH value 4.4, ultrasonic frequency 24kHz, ultrasonic reactor popped one's head in carry out ultrasonic enzymolysis (depth of penetration of popping one's head in immobilizes) in the insertion reaction liquid.At ultrasonic power is to stop ultrasonicly under the 30W behind the ultrasonic 10min, continues enzymolysis, and enzymolysis time is 4h.
(3) measure conversion rate of reducing sugar behind the enzymolysis at last: finish behind the enzymolysis the reaction solution centrifugation, supernatant liquor is surveyed its total reducing sugars content with the DNS method.Calculating conversion rate of reducing sugar by formula is 8.45%, than the raising 24.1% that does not add the auxiliary stalk cellulose hydrolysis of ultrasonic wave.
Embodiment six:
(1) feedstock production: wheat stalk is behind natural air drying, and it is standby to be crushed to 40 orders.
(2) the auxiliary stalk cellulose hydrolysis of ultrasonic wave: under the condition of concentration of substrate 50g/L, 50 ℃ of hydrolysis temperatures, pH value 5.2, ultrasonic frequency 24kHz, ultrasonic reactor popped one's head in carry out ultrasonic enzymolysis (depth of penetration of popping one's head in immobilizes) in the insertion reaction liquid.At ultrasonic power is to stop ultrasonicly under the 30W behind the ultrasonic 10min, continues enzymolysis, and enzymolysis time is 4h.
(3) measure conversion rate of reducing sugar behind the enzymolysis at last: finish behind the enzymolysis the reaction solution centrifugation, supernatant liquor is surveyed its total reducing sugars content with the DNS method.Calculating conversion rate of reducing sugar by formula is 8.65%, than the raising 27.2% that does not add the auxiliary stalk cellulose hydrolysis of ultrasonic wave.
Embodiment seven:
(1) feedstock production: wheat stalk is behind natural air drying, and it is standby to be crushed to 40 orders.
(2) the auxiliary stalk cellulose hydrolysis of ultrasonic wave: under the condition of concentration of substrate 50g/L, 50 ℃ of hydrolysis temperatures, pH value 4.8, ultrasonic frequency 24kHz, ultrasonic reactor popped one's head in carry out ultrasonic enzymolysis (depth of penetration of popping one's head in immobilizes) in the insertion reaction liquid.At ultrasonic power is to stop ultrasonicly under the 30W behind the ultrasonic 30min, continues enzymolysis, and enzymolysis time is 4h.
(3) measure conversion rate of reducing sugar behind the enzymolysis at last: finish behind the enzymolysis the reaction solution centrifugation, supernatant liquor is surveyed its total reducing sugars content with the DNS method.Calculating conversion rate of reducing sugar by formula is 16.46%, than the raising 142% that does not add the auxiliary stalk cellulose hydrolysis of ultrasonic wave.
Claims (1)
1. the method for the plain enzymolysis process of ultrasonic wave auxiliary fiber comprises the steps:
(1) feedstock production: wheat stalk is behind natural air drying, and it is standby to be crushed to 40 orders;
(2) the auxiliary stalk cellulose hydrolysis of ultrasonic wave: under concentration of substrate 50g/L, composite plant lytic enzyme concentration 0.5fbg/g, hydrolysis temperature 35-60 ℃, the condition of pH value 4.4-5.2, ultrasonic frequency 24kHz, ultrasonic reactor popped one's head in carry out ultrasonic assistance enzymolysis in the insertion reaction liquid, the depth of penetration of wherein popping one's head in immobilizes; At ultrasonic power is to stop ultrasonicly under the 0-60W behind the ultrasonic 0-40min, continues enzymolysis, and enzymolysis time is 4h;
(3) behind the enzymolysis with the reaction solution centrifugation, supernatant liquor.
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Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103343144A (en) * | 2013-07-26 | 2013-10-09 | 淮阴师范学院 | Method for pretreating to prepare bioethanol by hydrolysis of hydrochloric acid and enzymolysis of cellulose under ultrasonic waves |
| CN105029451A (en) * | 2015-06-30 | 2015-11-11 | 江苏大学 | Garlic straw active dietary fiber and preparation method thereof |
| CN105124711A (en) * | 2015-06-30 | 2015-12-09 | 江苏大学 | Preparation method of Chinese arrowhead fermented drinks |
| CN106538999A (en) * | 2016-11-25 | 2017-03-29 | 湖南宾之郎食品科技有限公司 | A kind of softening method of edible areca-nut |
| CN110074361A (en) * | 2018-01-26 | 2019-08-02 | 江苏省农业科学院 | A method of complex enzyme vigor and preparation high-quality strawberry fruit powder are improved by low-energy ultrasound |
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| CN101020720A (en) * | 2006-12-28 | 2007-08-22 | 宋桃森 | Process of extracting pueraria polysaccharide |
| CN101574153A (en) * | 2009-04-13 | 2009-11-11 | 宁夏中宁县早康枸杞开发有限公司 | Production method of liquid medlar |
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Cited By (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103343144A (en) * | 2013-07-26 | 2013-10-09 | 淮阴师范学院 | Method for pretreating to prepare bioethanol by hydrolysis of hydrochloric acid and enzymolysis of cellulose under ultrasonic waves |
| CN105029451A (en) * | 2015-06-30 | 2015-11-11 | 江苏大学 | Garlic straw active dietary fiber and preparation method thereof |
| CN105124711A (en) * | 2015-06-30 | 2015-12-09 | 江苏大学 | Preparation method of Chinese arrowhead fermented drinks |
| CN105029451B (en) * | 2015-06-30 | 2018-04-03 | 江苏大学 | A kind of garlic stalk activity dietary fiber and preparation method thereof |
| CN105124711B (en) * | 2015-06-30 | 2018-05-25 | 江苏大学 | A kind of preparation method of arrowhead fermented beverage |
| CN106538999A (en) * | 2016-11-25 | 2017-03-29 | 湖南宾之郎食品科技有限公司 | A kind of softening method of edible areca-nut |
| CN110074361A (en) * | 2018-01-26 | 2019-08-02 | 江苏省农业科学院 | A method of complex enzyme vigor and preparation high-quality strawberry fruit powder are improved by low-energy ultrasound |
| CN110074361B (en) * | 2018-01-26 | 2023-06-16 | 江苏省农业科学院 | Method for improving activity of compound enzyme and preparing high-quality strawberry fruit powder through low-intensity ultrasonic waves |
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Application publication date: 20110223 |