阅读说明：本技术 污水处理用辅助酶制剂制备方法 (Preparation method of auxiliary enzyme preparation for sewage treatment ) 是由 王林 罗学才 谷中华 尹若春 于 2019-10-09 设计创作，主要内容包括：污水处理用辅助酶制剂制备方法,所述方法包括：对漆酶、辣根过氧化物酶、化学模拟酶单独进行配置,并分别检测出各个酶活力的实际值；判断所述漆酶、辣根过氧化物酶、化学模拟酶活力的实际值是否达到预设值；若否,则重新配置；若是,则进行以下操作,包括：计算所述各个酶活力的实际值的增长率；将计算的所述增长率较大的两个酶进行混合,形成预制剂；将所述增长率最小的酶与所述预制剂进行混合,得到辅助酶制剂。本发明采用漆酶、辣根过氧化物酶、化学模拟酶采用特殊的混合方式根据酶的活力先后进行混合,确保辅助酶制剂能保证最高的活力进行污水处理,处理污水的效率高,同时无任何化学污染产生；对环境的要求低,操作简单,适合大规模的污水处理。(The preparation method of the auxiliary enzyme preparation for sewage treatment comprises the following steps: independently configuring laccase, horseradish peroxidase and chemical mimic enzyme, and respectively detecting actual values of enzyme activities; judging whether the actual values of the activities of the laccase, the horseradish peroxidase and the chemical simulation enzyme reach preset values or not; if not, reconfiguring; if yes, the following operations are carried out, including: calculating the growth rate of the actual value of each enzyme activity; mixing the two enzymes with the calculated larger growth rates to form a pre-formulation; mixing the enzyme with the minimum growth rate with the pre-formulation to obtain an auxiliary enzyme preparation. According to the invention, laccase, horseradish peroxidase and chemical mimic enzyme are mixed in sequence by adopting a special mixing mode according to the activity of the enzymes, so that the auxiliary enzyme preparation can ensure the highest activity to carry out sewage treatment, the sewage treatment efficiency is high, and no chemical pollution is generated; low requirement on environment, simple operation and suitability for large-scale sewage treatment.)

1. The preparation method of the auxiliary enzyme preparation for sewage treatment is characterized by comprising the following steps: the method comprises the following steps:

independently configuring laccase, horseradish peroxidase and chemical mimic enzyme, and respectively detecting actual values of enzyme activities;

judging whether the actual values of the activities of the laccase, the horseradish peroxidase and the chemical simulation enzyme reach preset values or not;

if not, reconfiguring;

if yes, the following operations are carried out, including:

calculating the growth rate of the actual value of each enzyme activity;

mixing the two enzymes with the calculated larger growth rates to form a pre-formulation;

mixing the enzyme with the minimum growth rate with the pre-formulation to obtain an auxiliary enzyme preparation.

2. The method for preparing an auxiliary enzyme preparation for sewage treatment according to claim 1, wherein: the mixing of the two enzymes whose growth rates are calculated to be larger includes:

adding the two enzymes with large growth rates at a constant speed in a vacuum tank with the temperature of 25-35 ℃ and the pH value of 3-5, and stirring at 2.0-2.4 r/min while adding;

stopping stirring after the two enzymes with larger growth rates are added, and standing for 5-10 min;

and (3) applying pressure to the vacuum tank, and continuously stirring for 10-20min at the speed of 1.0-1.8 r/min.

3. The method for preparing an auxiliary enzyme preparation for sewage treatment according to claim 1, wherein: said mixing the enzyme with the least growth rate with the pre-formulation comprises:

adding the enzyme with the minimum growth rate at a constant speed in a vacuum tank with the temperature of 25-35 ℃ and the pH value of 3-5, and stirring at 3.2-4.3 r/min while adding;

stopping stirring after the enzyme with the minimum growth rate is added, and standing for 10-16 min;

and (3) applying pressure to the vacuum tank, and continuously stirring for 10-20min at the speed of 1.8-2.4 r/min.

4. The process for preparing an auxiliary enzyme preparation for sewage treatment according to any one of claims 1 to 3, wherein: the configuration method of the chemical mimic enzyme comprises the following steps:

stirring S-adenosylmethionine and copper chloride at room temperature and concentration of 20mmol/L and pH of 5.5 in acetic acid-sodium acetate buffer solution and molar ratio of 2:3 for full reaction;

and centrifuging the reacted reaction solution at 8000rpm for 5min, and taking the supernatant to obtain the chemical mimic enzyme.

5. The process for preparing an auxiliary enzyme preparation for sewage treatment according to any one of claims 1 to 3, wherein: the preset values of the enzyme activities of the laccase, the horseradish peroxidase and the chemical simulation enzyme are respectively as follows: 2000u/ml, 3000u/ml and 500 u/ml.

6. The process for preparing an auxiliary enzyme preparation for sewage treatment according to any one of claims 1 to 3, wherein: the calculation formula for calculating the increase rate of the actual value of each enzyme activity comprises the following steps:

wherein An represents the increase rate of the nth enzyme, wherein laccase is defined as n ═ 1, horseradish peroxidase is defined as n ═ 2, chemomimetic enzyme activity is defined as n ═ 3, and X is defined as_{n actual}Is the actual activity value of the nth enzyme, X_{n is preset}The preset activity value for the nth enzyme.

7. The process for preparing an auxiliary enzyme preparation for sewage treatment according to any one of claims 1 to 3, wherein: the addition ratio of the laccase to the horseradish peroxidase to the chemical mimic enzyme is 2.2: 1.8: 1.

8. the process for preparing an auxiliary enzyme preparation for sewage treatment according to any one of claims 1 to 3, wherein: the activity of the laccase in the auxiliary enzyme preparation is more than or equal to 500u/ml, the activity of the horseradish peroxidase is more than or equal to 300u/ml, and the activity of the chemical mimic enzyme is more than or equal to 50 u/ml.

9. The process for preparing an auxiliary enzyme preparation for sewage treatment according to any one of claims 1 to 3, wherein: and the step of detecting the actual value of each enzyme activity comprises the step of detecting the actual value of the enzyme activity by adopting an ultraviolet spectrophotometer.

Technical Field

The invention belongs to the technical field of sewage treatment, and particularly relates to a preparation method of an auxiliary enzyme preparation for sewage treatment.

Background

At present, sewage plants have many problems in the operation process, such as: the effluent quality is unstable, and the shock resistance is poor; the effluent is also disinfected by ultraviolet rays or liquid chlorine or other means, so that the water body loses self-cleaning capability due to the death of microorganisms; the root cause of these problems is the insufficient decomposition capacity of microorganisms in sewage plants; in order to improve the decomposition ability, enzyme preparations are cited; the enzyme preparation is a biological product with a catalytic function after purification and processing of enzyme, is mainly used for catalyzing various chemical reactions in the production process, and has the characteristics of high catalytic efficiency, high specificity, mild action conditions, energy consumption reduction, chemical pollution reduction and the like after being used for sewage treatment.

However, most of the existing enzyme preparations are single enzyme preparations, the catalytic effect of single enzyme is limited due to the fact that organic matters in sewage are abundant and disordered, and when the sewage is treated by simply mixing a plurality of enzymes, the activity of the enzyme is reduced due to the mixing of the enzymes, so that the sewage treatment by simply mixing the enzymes at the highest activity can not be guaranteed, and the efficiency is low.

Disclosure of Invention

Aiming at the defects in the prior art, the invention provides a preparation method of an auxiliary enzyme preparation for sewage treatment, and the specific technical scheme is as follows:

independently configuring laccase, horseradish peroxidase and chemical mimic enzyme, and respectively detecting actual values of enzyme activities;

judging whether the actual values of the activities of the laccase, the horseradish peroxidase and the chemical simulation enzyme reach preset values or not;

if not, reconfiguring;

if yes, the following operations are carried out, including:

calculating the growth rate of the actual value of each enzyme activity;

mixing the two enzymes with the calculated larger growth rates to form a pre-formulation;

mixing the enzyme with the minimum growth rate with the pre-formulation to obtain an auxiliary enzyme preparation.

Further, the mixing the two enzymes having the calculated larger growth rates includes:

adding the two enzymes with large growth rates at a constant speed in a vacuum tank with the temperature of 25-35 ℃ and the pH value of 3-5, and stirring at 2.0-2.4 r/min while adding;

stopping stirring after the two enzymes with larger growth rates are added, and standing for 5-10 min;

and (3) applying pressure to the vacuum tank, and continuously stirring for 10-20min at the speed of 1.0-1.8 r/min.

Further, said mixing said minimally-increasing enzyme with said pre-formulation comprises:

adding the enzyme with the minimum growth rate at a constant speed in a vacuum tank with the temperature of 25-35 ℃ and the pH value of 3-5, and stirring at 3.2-4.3 r/min while adding;

stopping stirring after the enzyme with the minimum growth rate is added, and standing for 10-16 min;

and (3) applying pressure to the vacuum tank, and continuously stirring for 10-20min at the speed of 1.8-2.4 r/min.

Further, the configuration method of the chemical mimic enzyme comprises the following steps:

stirring S-adenosylmethionine and copper chloride at room temperature and concentration of 20mmol/L and pH of 5.5 in acetic acid-sodium acetate buffer solution and molar ratio of 2:3 for full reaction;

and centrifuging the reacted reaction solution at 8000rpm for 5min, and taking the supernatant to obtain the chemical mimic enzyme.

Further, the preset values of the activities of the laccase, the horseradish peroxidase and the chemical simulation enzyme are respectively as follows: 2000u/ml, 3000u/ml and 500 u/ml.

Further, the calculation formula for calculating the increase rate of the actual value of each enzyme activity includes:

in the formula, An represents the growth rate of the nth enzyme, laccase is used when n is 1, horseradish peroxidase is used when n is 2, chemomimetic enzyme activity is used when n is 3, Xn is actually the actual activity value of the nth enzyme, and Xn is preset to be the preset activity value of the nth enzyme.

Further, the addition ratio of the laccase, the horseradish peroxidase and the chemical mimic enzyme is 2.2: 1.8: 1.

furthermore, the activity of the laccase in the auxiliary enzyme preparation is more than or equal to 500u/ml, the activity of the horseradish peroxidase is more than or equal to 300u/ml, and the activity of the chemical mimic enzyme is more than or equal to 50 u/ml.

Further, the step of detecting the actual value of each enzyme activity comprises detecting the actual value of the enzyme activity by using an ultraviolet spectrophotometer.

The invention has the beneficial effects that: according to the invention, laccase, horseradish peroxidase and chemical mimic enzyme are mixed in sequence by adopting a special mixing mode according to the increase rate of enzyme activity, so that the auxiliary enzyme preparation can ensure the highest activity to carry out sewage treatment, the sewage treatment efficiency is high, and no chemical pollution is generated; low requirement on environment, simple operation and suitability for large-scale sewage treatment.

Drawings

FIG. 1 shows a schematic flow chart of a preparation method of an auxiliary enzyme preparation for sewage treatment according to an embodiment of the invention.

Detailed Description

In order to make the objects, technical solutions and advantages of the present invention more apparent, the present invention is further described in detail with reference to the following embodiments. It should be understood that the specific embodiments described herein are merely illustrative of the invention and are not intended to limit the invention.

The invention provides a preparation method of an auxiliary enzyme preparation for sewage treatment, which comprises the following steps, as shown in figure 1 by way of example:

the method comprises the following steps: independently configuring laccase, horseradish peroxidase and chemical mimic enzyme, and respectively detecting actual values of enzyme activities;

specifically, aspergillus oryzae is adopted as a strain for producing the laccase, horseradish is adopted as a strain for producing the horseradish peroxidase, and the chemical mimic enzyme refers to an enzyme amount which is equivalent to that required for converting 1uM catechol per minute under the condition that the pH is 5.0 and the temperature is 25 ℃, wherein the preparation method of the chemical mimic enzyme comprises the steps of stirring and fully reacting at room temperature, wherein the concentration of the enzyme amount is 20mmol/L, the pH is 5.5, acetic acid-sodium acetate buffer solution, and S-adenosylmethionine and copper chloride are used in a molar ratio of 2: 3; and centrifuging the reacted reaction solution at 8000rpm for 5min, and taking the supernatant to obtain the chemical mimic enzyme.

The addition ratio of the laccase, the horseradish peroxidase and the chemical mimic enzyme is 2.2: 1.8: 1. the actual value of each enzyme activity detected in the invention includes the actual value of the enzyme activity detected by an ultraviolet spectrophotometer, but is not limited thereto.

Step two: judging whether the actual values of the activities of the laccase, the horseradish peroxidase and the chemical simulation enzyme reach preset values or not;

if not, reconfiguring;

if yes, the following operations are carried out, including:

calculating the growth rate of the actual value of each enzyme activity;

mixing the two enzymes with the calculated larger growth rates to form a pre-formulation;

mixing the enzyme with the minimum growth rate with the pre-formulation to obtain an auxiliary enzyme preparation.

Specifically, the invention limits the preset values of the activities of the laccase, the horseradish peroxidase and the chemical simulation enzyme to be respectively as follows: 2000u/ml, 3000u/ml and 500 u/ml. Comparing the preset value with the actual value detected in the first step, judging whether the actual value is greater than or equal to the preset value, and if not, reconfiguring the corresponding enzyme preparation; if so, indicating that the enzyme per se achieves the corresponding activity, performing the following operations, including:

2.1 calculating the growth rate of the actual value of each enzyme activity;

specifically, the calculation formula for calculating the increase rate of the actual value of each enzyme activity includes:

wherein An represents the increase rate of the nth enzyme, wherein laccase is defined as n ═ 1, horseradish peroxidase is defined as n ═ 2, chemomimetic enzyme activity is defined as n ═ 3, and X is defined as_{n actual}Is the actual activity value of the nth enzyme, X_{n is preset}The preset activity value for the nth enzyme.

2.2 mixing the two enzymes with larger calculated growth rate to form a pre-preparation;

specifically, after the increase rate calculation is completed, the values of the activity increase rates of the three enzymes are compared, the two enzymes with the larger increase rates are premixed firstly, then the enzyme with the smallest increase rate is mixed, and the activity of the enzyme is reduced in the later mixing, so that the enzyme with the higher activity increase rate is mixed firstly, and finally the enzyme with the smallest activity is processed, so that the overall activity of the auxiliary enzyme preparation can be effectively improved.

Specifically, the mixing the two enzymes with the calculated larger growth rates comprises:

adding the two enzymes with large growth rates at a constant speed in a vacuum tank with the temperature of 25-35 ℃ and the pH value of 3-5, and stirring at 2.0-2.4 r/min while adding;

stopping stirring after the two enzymes with larger growth rates are added, and standing for 5-10 min;

and (3) applying pressure to the vacuum tank, and continuously stirring for 10-20min at the speed of 1.0-1.8 r/min. The two materials are quickly mixed by stirring at a higher stirring speed, then are kept stand to be melted, and finally are stirred again to ensure the complete mixing.

2.3 mixing the enzyme with the minimum growth rate with the pre-preparation to obtain an auxiliary enzyme preparation;

specifically, said mixing said minimally-increasing enzyme with said pre-formulation comprises:

adding the enzyme with the minimum growth rate at a constant speed in a vacuum tank with the temperature of 25-35 ℃ and the pH value of 3-5, and stirring at 3.2-4.3 r/min while adding;

stopping stirring after the enzyme with the minimum growth rate is added, and standing for 10-16 min;

and (3) applying pressure to the vacuum tank, and continuously stirring for 10-20min at the speed of 1.8-2.4 r/min. The two materials are quickly mixed by stirring at a higher stirring speed, then are kept stand to be melted, and finally are stirred again to ensure the complete mixing. The activity of the laccase in the mixed auxiliary enzyme preparation is more than or equal to 500u/ml, the activity of the horseradish peroxidase is more than or equal to 300u/ml, and the activity of the chemical mimic enzyme is more than or equal to 50 u/ml.