Method for removing adhesive in waste paper pulp by using composite enzyme preparation

2024-04-12 20:06:50

Patent Publication No.: CN 106318930 A

Applicant: South China University of Technology

Inventor: Wang Yonghua, Lan Dongming, Zhang Zedong, Yang Bo, Zhou Pengfei

A method for removing the adhesive in waste paper pulp by a composite enzyme preparation mainly composed of high temperature esterase TTEST is provided.

Proceed as follows:

(1) Preparation of high temperature esterase TTEST:

The esterase TTEST strain was inoculated into the seed liquid medium to shake culture, and then the strain was inoculated into the fermentation medium to shake culture, and the high temperature esterase TTEST crude enzyme solution was prepared, and the high temperature esterase TTEST crude enzyme solution was purified. The enzymatic activity of the high temperature esterase TTEST was determined by phenol colorimetry.

The gene encoding the TTEST esterase obtained by the whole gene synthesis method was cloned into the expression vector pET23a-CBD vector to obtain a recombinant expression vector. The expression plasmid containing the TTEST esterase-encoding gene was transferred into the BL21 (DE3) strain by the CaCl2 transformation method to obtain a genetically engineered strain. The strain of esterase TTEST was introduced into LB seed medium containing Amp (100 ug/ml) to expand the culture. When the OD value reached 0.6-0.8, the bacterial solution was inoculated to Amp-containing (100 ug/r) at a ratio of 1:100. In the LB fermentation medium of ml), the expression of I PTG (20 mmol/L) inducer was induced by shaking until the OD value was 0.6-0.8, and the cells were collected after 18-24 hours. After the cells were centrifuged at 12000 r/min for 10 min, the cells of the tube wall were collected for sonication, and then centrifuged at 12000 r/min for 10 min, and then the supernatant was taken to obtain a crude enzyme solution. Specific binding of cellulose to esterase is carried out, followed by digestion with 3C protease to obtain a pure enzyme. 1L of bacterial liquid corresponds to 1g of cellulose, and the crude enzyme solution is combined with cellulose at room temperature for 30 minutes, and stirred at the appropriate time. After combining, the cells were centrifuged at high speed for 10 min, the supernatant was discarded, and the adsorbed protein was washed twice with PBS buffer. The 3C protease was centrifuged before use, the supernatant was discarded, and washed twice with PBS buffer. An appropriate amount of PBS was added to the adsorbed protein, and 1 L of the bacterial solution was mixed with 1 ml of 3C protease, and then digested for 4 hours or overnight. After centrifugation, the cells are centrifuged at high speed, and the supernatant is pure enzyme. Thereafter, the enzyme activity of the prepared high temperature esterase TTEST was measured by a p-nitrophenol colorimetric method, and the enzyme activity was found to be 3500 U/g. The lipase CALB is a commercial lipase, and the enzyme activity was measured to be 150.0 U/ml.

(2) According to the mass percentage, high temperature esterase TTEST (45% ~ 50%) and amylase (15% ~ 20%), pectinase (10% ~ 15%), xylanase (10% ~ 15%), glycerin (5%), polyethylene glycol (5%), and water (5% to 10%) are compounded into a composite enzyme preparation.

(3) Weigh 100g OCC absolute dry pulp, dilute to the corresponding pulp concentration, add a certain amount of enzyme, at a certain processing temperature, pH value, treatment time, stirring speed (as shown in Table 1), according to TAPPIT-277 adhesive The slurry was screened by a Pulmac-Master Screen sifter, and then the sifted gel was dyed, compressed, and finally scanned and scanned by a scanning software to determine the content of the adhesive after the enzyme treatment.

The waste paper pulp was treated under the same conditions without any enzyme, and the experimental results were the blank group adhesive content.

The compound enzyme preparation is added in an amount of 0.1% to 1.0% by mass of the waste pulp slurry, a treatment temperature of 40 to 80 ° C, a treatment time of 45 to 120 minutes, a pH of 6.0 to 9.0, and a stirring speed of 100 to 150 r/min.

The temperature of the composite enzyme treated waste paper pulp is 50-55 ° C, the stirring condition of the composite enzyme-treated waste paper pulp is 120 r / min, and the reaction time is 60 min. The amount of the complex enzyme added is 0.3% to 0.5% of the mass of the waste paper pulp. The pH of the complex enzyme-treated waste paper pulp is 7.0 to 8.0. The pulp of waste paper pulp is 4% to 6%.

The enzyme activity of the high temperature esterase was 3500 U/g. The amylase activity was 5000 U/g, the pectinase activity was 5000 U/g, and the xylanase activity was 10000 U/g.


It can be seen from Table 1 that under the condition of 40-55 °C, the content of the adhesive in the waste paper pulp treated with the compound enzyme is obviously reduced, and the removal rate of the adhesive can reach 60.8%-74.1%, which is obviously superior to the lipase CALB. Removal. Even under the high temperature condition of 80 °C, the treatment effect after adding the complex enzyme is ideal, and the adhesive removal rate in the waste paper pulp reaches 58.5%, and the lipase CALB is basically inactivated under this condition, and it is difficult to remove the adhesive. Considering that the temperature of different sections of the waste papermaking process is not the same, the esterase PCEST is ideal for removing the adhesive in different sections of waste paper recycling.

Benefits compared to the prior art:

(1) The preparation of high temperature esterase TTEST is relatively easy and the effect is obvious.

(2) The composite enzyme preparation can cope with various high temperature and acid-base environments in the waste paper recycling process, and can still maintain good adhesive removal effect under high temperature conditions.

(3) The method of removing the waste paper glue by the high-temperature esterase-based composite enzyme preparation can be directly carried out on the production line without adding any mechanical equipment, and the operation is simple and the cost is low.

Therefore, the method for controlling the adhesive in the waste paper pulp by the high-temperature esterase-based composite enzyme preparation efficiently and steadily solves the problem that the product quality in the waste papermaking field seriously affects the product quality in the past, and has the advantages of simple operation and removal effect. Significant, low production cost, no pollution and so on.

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