阅读说明：本技术 一种高浓度、低分子量液体壳寡糖的生产工艺 (Production process of high-concentration low-molecular-weight liquid chitosan oligosaccharide ) 是由 张健 苏国金 王昕宇 于 2020-05-07 设计创作，主要内容包括：本发明公开了一种高浓度、低分子量液体壳寡糖的生产工艺,属于生物工程技术领域,解决了高浓度液体壳寡糖生产困难、高分子产品不易吸收、混配性差的问题。其技术要点是：高浓度壳聚糖加酸溶胀,反应釜中增加通风,辅助搅拌均匀,停风、停搅拌静置2-5h,然后加入过量特异性高酶活壳聚糖酶进行酶解反应,同时进行通风,待粘度降至80mpa.s-100mpa.s时,开启搅拌,停止通风,搅拌至反应结束。该工艺生产得到的液体壳寡糖含量可达到20-25％,远超过现有市场上壳寡糖产品的正常生产浓度,分子量可控制在600-1500Da之间,被植物吸收利用后能更好的发挥其诱抗作用。该发明可大大降低生产能耗,节约生产成本,同时也降低了运输费用等,更好的提高了生产效率。(The invention discloses a production process of high-concentration low-molecular-weight liquid chitosan oligosaccharide, belongs to the technical field of bioengineering, and solves the problems of difficult production of high-concentration liquid chitosan oligosaccharide, difficult absorption of high-molecular products and poor mixing property. The technical key points are as follows: adding acid to swell the high-concentration chitosan, adding ventilation in a reaction kettle, assisting in stirring uniformly, stopping ventilation, stopping stirring, standing for 2-5h, then adding excessive specific high-enzyme activity chitosanase to perform enzymolysis reaction, simultaneously performing ventilation, starting stirring when the viscosity is reduced to 80-100mpa.s, stopping ventilation, and stirring until the reaction is finished. The content of the liquid chitosan oligosaccharide produced by the process can reach 20-25%, which is far higher than the normal production concentration of chitosan oligosaccharide products in the existing market, the molecular weight can be controlled between 600-1500Da, and the induced resistance effect can be better played after the chitosan oligosaccharide is absorbed and utilized by plants. The invention can greatly reduce the production energy consumption, save the production cost, simultaneously reduce the transportation cost and the like, and better improve the production efficiency.)

1. A production process of high-concentration low-molecular-weight liquid chitosan oligosaccharide is characterized by comprising the following steps: adding acid to swell the high-concentration chitosan, adding ventilation in a reaction kettle, uniformly stirring in an auxiliary way, stopping wind, stopping stirring, standing for 2-5h, adding excessive specific high-enzyme activity chitosanase after swelling is finished, carrying out enzymolysis reaction, simultaneously ventilating, starting stirring when the viscosity is reduced to 80-100mpa.s, stopping ventilation, and stirring until the reaction is finished.

2. The process for producing high-concentration, low-molecular-weight liquid chitosan oligosaccharide according to claim 1, wherein: the highest adding concentration of the raw material chitosan can reach 25 percent, the chitosan is flaky or powdery solid, the chitosan is added into water and then stirred uniformly, and the temperature is controlled at 35-45 ℃.

3. The process for producing high-concentration, low-molecular-weight liquid chitosan oligosaccharide according to claim 1, wherein: the acid used in the swelling process is one or the mixture of any of formic acid, acetic acid, lactic acid, hydrochloric acid, nitric acid and citric acid.

4. The process for producing high-concentration, low-molecular-weight liquid chitosan oligosaccharide according to claim 1, wherein: the reaction kettle is ventilated, and the ventilation quantity is controlled to be more than or equal to 0.5 vvm.

5. The process for producing high-concentration, low-molecular-weight liquid chitosan oligosaccharide according to claim 1, wherein: the enzyme activity of the chitosanase is more than or equal to 2000U, the addition amount of the chitosanase is 2-5 times of the normal dosage, the reaction temperature is 35-45 ℃, the reaction time is 2-6h, and the ventilation volume is more than or equal to 1 vvm.

6. The process for producing high-concentration, low-molecular-weight liquid chitosan oligosaccharide according to claim 1, wherein: the end of the reaction is marked by the viscosity of the reaction solution decreasing to 3.8-4.0mpa.s, and the molecular weight can be controlled between 600 and 1500 Da.

Technical Field

The invention relates to the technical field of bioengineering, in particular to a production process of high-concentration low-molecular-weight liquid chitosan oligosaccharide.

Background

At present, the products of the chitosan oligosaccharide are widely applied to various industries such as agriculture, medicine, health care and the like. At present, the production of chitosan oligosaccharide is mainly prepared by a biological enzyme method, but is limited by the viscosity of raw material chitosan, so that the content of a chitosan oligosaccharide liquid product is always kept at a relatively low level. It is reported that the highest content of liquid chitosan oligosaccharide production is about 15%, which increases the cost in transportation and subsequent spray concentration.

As for the molecular weight of chitosan oligosaccharide, the number average molecular weight of most products is controlled between 1500-4000Da at present, and the high molecular weight (the molecular weight is more than or equal to 5000Da) is relatively high. There are two major problems with relatively high molecular weight chitosan oligosaccharide products: firstly, the plant is not easy to absorb and utilize; secondly, the mixing property with other preparations is poor. It is necessary to control the low molecular weight products.

The production of the high-concentration low-molecular-weight liquid chitosan oligosaccharide can reduce the production and transportation cost, save the energy consumption, labor and the like of later spray drying, save the product cost to a greater extent, and simultaneously, the low-molecular-weight product not only is more beneficial to the absorption and utilization of plants, but also can be better compounded with other preparation products for use, thereby enhancing the application effect. Therefore, it is desirable to provide a process for producing high-content, low-molecular-weight chitosan oligosaccharides.

Disclosure of Invention

Aiming at the problems, the invention provides a production process of high-concentration low-molecular-weight liquid chitosan oligosaccharide, which is mainly characterized in that the process is improved on the basis of the original production of chitosan oligosaccharide by biological enzymolysis, the content of the finally obtained liquid chitosan oligosaccharide product is 20-25%, and the molecular weight of the product is 600-1500 Da.

The specific process technology comprises the following steps: adding acid to swell the high-concentration chitosan, adding ventilation in a reaction kettle, assisting in stirring uniformly, stopping ventilation, stopping stirring, standing for 2-5h, then adding excessive specific high-enzyme activity chitosanase to perform enzymolysis reaction, simultaneously performing ventilation, starting stirring when the viscosity is reduced to 80-100mpa.s, stopping ventilation, and stirring until the reaction is finished.

Wherein the chitosan is flake or powdery solid, and is added into water and uniformly stirred, and the temperature is controlled at 35-45 ℃; the acid used in the swelling process is one or the mixture of any more of formic acid, acetic acid, lactic acid, hydrochloric acid, nitric acid and citric acid; the air quantity for increasing ventilation in the reaction kettle is controlled to be more than or equal to 0.5 vvm; the enzyme activity of the chitosanase used in the process is more than or equal to 2000U, the addition amount is 2-5 times of the normal dosage, the reaction temperature is 35-45 ℃, the reaction time is 2-6h, and the ventilation volume is more than or equal to 1 vvm; the end of the reaction was judged as a decrease in the viscosity of the reaction solution to 3.8-4.0 mpa.s.

The content of the liquid chitosan oligosaccharide produced by the process can reach 20-25%, which is far higher than the normal production concentration of chitosan oligosaccharide products in the existing market, the molecular weight can be controlled between 600-1500Da, and the induced resistance effect can be better played after the chitosan oligosaccharide is absorbed and utilized by plants. The invention can greatly reduce the production energy consumption, save the production cost, simultaneously reduce the transportation cost and the like, and better improve the production efficiency.

Detailed Description

The present invention will be described in further detail with reference to the following examples, but it should not be construed that the scope of the above subject matter is limited to the following examples. All the technologies realized based on the above contents of the present invention belong to the scope of the present invention.