CN100478448C - Enzyme catalysis method for continuous synthesis of lauroyl maltose under organic phase - Google Patents

Abstract

The invention discloses a continuous synthesizing method of maltose laurate in the organic phase, which is characterized by the following: using stirring tank typed enzyme reactor to synthesize maltose laurate; affirming optimum manufacturing condition; maintaining the reactor production at 10g/(L .d) for 8-10d; obtaining the purity of product to 95%.

Description

A kind of method that the maltose laurate is synthesized in enzyme catalysis continuously in organic phase

Technical field

The present invention relates to a kind of in organic phase the lipase-catalyzed continuously method of synthetic nonionic surface active agent maltose laurate, belong to the biological food chemical technology field.

Background technology

Fatty ester of maltose is the edible tensio-active agent with property of a class that is formed by maltose and lipid acid condensation.Because fatty ester of maltose has good emulsifying, viscous resistance and puffing and can be widely used in grocery trade, cosmetic industry and washing Industry.In recent years, abroad some investigators increase gradually to the enzyme process synthesis technique of fatty ester of maltose and the research that is applied in the food.

In China the report of the synthetic fatty ester of maltose of enzyme process catalysis in organic phase is not arranged as yet, but the synthetic report of enzyme process of some other sugar esters is arranged, people such as Zhang Nianxiang reported the fatty acid ester of lipase-catalyzed synthetic different monose in organic phase in 96 years.Feng Leigang and Guozheng ZHANG have been reported the technical study of synthesizing amylaceum laurate in the nonaqueous phase in 2004.Wu Keke and Anqing big (2004) are raw material with sucrose and palmitinic acid, have studied its enzyme process synthetic technology.But these reports only are confined to the research of batch production technology.

At present commercially available sugar ester product is chemosynthesis basically, and reaction is to carry out under high temperature, high pressure, and product be the mixture of complexity, obtain highly purified product and need carry out numerous and diverse mask work, and product often has darker color and luster.And adopt the batch reactor enzyme process synthetic, the solubleness of substrate maltose in reaction system acetone only is 0.59mmol/L, speed of reaction is slower, and needs in the reaction to add molecular sieve as dewatering agent, thus and because product is accumulated inhibited reaction in reactor carrying out.Therefore, batch production is not suitable for large-scale industrial production.

Summary of the invention

The purpose of this invention is to provide a kind of method that the maltose laurate is synthesized in enzyme catalysis continuously in organic phase, by designing a kind of lipase-catalyzed continuous synthetic reactor that is fit to, provide the method for the synthetic nonionic surface active agent maltose laurate of industrialization.Simply be convenient to separation and purification with present method mild condition, reaction efficiency height, products therefrom composition, can realize industrial-scale production, obviously be better than chemical method and batch production.

Technical scheme of the present invention: utilizing continuously stirring pot type enzyme reactor, is raw material with maltose and lauric acid, and lipase is the synthetic maltose laurate of catalyzer, obtains product maltose laurate through separation and purification.Manufacturing condition is:

(1) the maltose initial concentration is 25-200mmol/L in the reactor;

(2) subsequently in the reaction process every day add maltose, addition is 8-20g/L;

(3) the lauric acid concentration in the feeding liquid lauric acid acetone soln is 100-400mmol/L;

(4) consumption of catalyzer lipase NOVE435 is 20-80g/L;

(5) adopt continuous reaction, in reaction process with constant flow pump with in the lauric acid acetone soln suction reactor, the reaction solution of equal volume is drawn out of simultaneously, constant flow pump flow velocity 0.08-0.36mL/min reacted 10-12 days;

(6) with the reaction solution underpressure distillation of collecting, reclaim solvent acetone, obtain solid matter simultaneously;

(7) solid that previous step is obtained adopts normal hexane repeatedly to wash purifying, and normal hexane consumption by volume after the product vacuum-drying that obtains after the washing, is maltose laurate product for 0.1-0.5 times of the reaction solution volume of collection.

Optimized process conditions is:

(1) the maltose initial concentration is 50mmol/L in the reactor;

(2) subsequently in the reaction process every day add maltose, addition is 16g/L;

(3) the lauric acid concentration in the feeding liquid lauric acid acetone soln is 200mmol/L;

(4) consumption of catalyzer lipase NOVE435 is 40g/L;

(5) adopt continuous reaction, in reaction process with constant flow pump with in the lauric acid acetone soln suction reactor, the reaction solution of equal volume is drawn out of simultaneously, constant flow pump flow velocity 0.15mL/min reacted 10-12 days;

(6) with the reaction solution underpressure distillation of collecting, reclaim solvent acetone, obtain solid matter simultaneously;

(7) solid that previous step is obtained adopts normal hexane washing purifying 3 times, and normal hexane consumption by volume is 0.25 times of the reaction solution volume collected for the first time, and the back is 0.1 times twice, after the product vacuum-drying that obtains after the washing, is maltose laurate product.

Isolate unreacted lauric acid simultaneously and continue to participate in reaction, evaporate organic solvent and recycle.The research of lipase-catalyzed maltose laurate method for continuously synthesizing comprises reactor that design is fit to, obtains maltose initial concentration and with the influence to output of the addition in the afterreaction, lauric acid concentration, lipase addition, flow velocity by experiment, determines the separation purification method and the continuous production processes flow process of product.

The design (Fig. 1) of the stirring pot type enzyme reactor of continuous feeding and discharging: reactor volume is 250mL, the iron basket of a band sieve aperture is fixed on the reactor cap, lipase is placed in one, and by the continuous stirring reaction liquid of magnetic stirring apparatus, there is water bath with thermostatic control control temperature of reaction system the outside.The lauric acid acetone soln enters in the reactor by constant flow pump, and effusive reaction solution is collected in constant current simultaneously.

Maltose initial concentration and determining: when the reaction beginning, add maltose concentration (mmol/L) and be respectively 25,50,100,150,200 with the addition in the afterreaction.Found that when the maltose initial concentration is too high, production concentration is reduced.When the maltose initial concentration is 50mmol/L, maltose ester concentration maximum in the reaction solution.Added 50mmol/L maltose in first day, and can find that two days later the undissolved maltose in the reactor was exhausted,, must add maltose in order to keep continuous production.Later on add 8-20g/L maltose every day in reactor, experimental result shows adds 16g/L every day, promptly can make to react to maintain under the higher production concentration during 4g maltose and carry out continuously.

Determining of lauric acid concentration: the concentration of lauric acid acetone soln (mmol/L) is respectively 100,200,300,400, selects for use 200mmol/L can obtain high yield.

Determining of catalyzer lipase NOVE435 addition: the addition of lipase is 20-80g/L, and addition is many more, and output is high more, when addition during greater than 40g/L, increase of production is slow, considers the price and the catalytic efficiency of enzyme, is 40g/L so lipase NOVE435 adds preferable amount.

The selection of reaction solution flow velocity: the reaction solution flow velocity is 0.08-0.36mL/min, changes flow velocity and promptly changes the residence time of lauric acid in reactor, if the residence time is too short, raw material is not fully reaction as yet, production concentration is lower, and the flow velocity that slows down, the prolongation residence time can make production concentration increase gradually.When slowing to certain flow rate, reaction solution can reach a steady state, and production concentration no longer increases.When charging flow velocity during greater than 1.5mL/min, the production concentration in the reaction solution descends gradually, and final flow rate preferably is defined as 1.5mL/min.

The separation purification method of reaction solution: reaction solution steams reaction solvent acetone by underpressure distillation, the normal hexane that adds different amounts subsequently dissolves unreacted lauric acid, obtain precipitation with the centrifugal 20min of the speed of 4000r/min, sedimentary main component is the maltose laurate.For the first time the ratio of the addition of normal hexane and former reaction solution is 1: 4, again with V _{Normal hexane}: V _{Reaction solution}=1: 10 addition handles twice, can obtain the product of ester content about 95%.Solution the inside main component after normal hexane is handled is unreacted lauric acid, steams normal hexane by underpressure distillation, and lipid acid continues to enter reactor reaction.The process flow sheet of successive reaction as shown in Figure 2.

Maltose laurate method for continuously synthesizing after the optimization of the present invention is: the concentration of maltose is 50mmol/L, lauric acid concentration is 200mmol/L, enzyme concentration is 40g/L, add 16g/L maltose every day, the lauric acid acetone soln of 200mmol/L enters reactor with the 0.15ml/min flow velocity, the output of maltose laurate can maintain greater than 10g/ (Ld) continuously in 8-10 days, utilized the lauric output that reclaims also can reach 10g/ (Ld).

Batch production and quantity-produced output and product are formed contrast: adopt the optimum process condition batch production, the concentration of maltose is 25mmol/L, the acid alcohol mol ratio is 3: 1, the molecular sieve addition is 80mg/mL, the lipase addition is 20mg/mL, 50 ℃ of shaking bath reaction 72h, the output of maltose laurate is about 4g/ (Ld), in the reaction product maltose lauric acid diester is arranged simultaneously.

Beneficial effect of the present invention: the present invention adopts the stirring enzyme reactor of continuous feeding and discharging, the method of continuously synthetic maltose laurate can improve the output of product and the efficient of reaction greatly, the output of reactor can reach about 13g/ (Ld), and does not need to add dewatering agent in the process of reaction.Owing to do not contain diester substantially in the reaction product, institute is so that separation and purification is fairly simple, and after 3 normal hexane washings, the retention rate of product reaches 80%, and the purity that obtains product simultaneously has 95%.The inventive method has and can amplify scale production, mild condition and reaction efficiency advantages of higher with industry.In the engineering of producing, because reaction is a reaction medium with the organic solvent, do not have the influence of microorganism in the production process substantially, and the solvent that is adopted is acetone and normal hexane, these two kinds all can be used in foodstuffs industry.The maltose laurate product that obtains can be used as tensio-active agent and is applied to grocery trade, cosmetic industry and washing Industry.

Description of drawings

The stirring enzyme reaction device of Fig. 1 continuous feeding and discharging.1. charging bottle, 2. constant flow pump, 3. magnetic stirring apparatus, 4. water-bath, 5. reactor, 6. the iron basket of band sieve aperture, 7. discharging bottle.

Fig. 2 continuous production processes flow process.

Embodiment

Embodiment 1

It is 25mmol/L (2.125g) that reaction is initially at the concentration of adding maltose in the reactor, 100mmol/L lauric acid acetone soln 250ml, 20g/L lipase, its afterreaction adds 8g/L maltose every day in reactor, the lauric acid acetone soln of 100mmol/L enters reactor with the 0.36ml/min flow velocity, reaction 10d.The product volume that obtains after output and the separation and purification sees Table 1.

Table 1

Embodiment 2

It is 50mmol/L (4.275g) that reaction is initially at the concentration of adding maltose in the reactor, 200mmol/L lauric acid acetone soln 250ml, 40g/L lipase, its afterreaction adds 12g/L maltose every day, the lauric acid acetone soln of 200mmol/L enters reactor with the 0.2ml/min flow velocity, reaction 10d.The product volume that obtains after output and the separation and purification sees Table 2.

Table 2

Embodiment 3

It is 100mmol/L that reaction is initially at the concentration of adding maltose in the reactor, 300mmol/L lauric acid acetone soln 250ml, 60g/L lipase, its afterreaction add 16g/L maltose every day, the lauric acid acetone soln of 300mmol/L enters reactor with the 0.15ml/min flow velocity, reaction 10d.The product volume that obtains after output and the separation and purification sees Table 3.

Table 3

Embodiment 4

It is 200mmol/L that reaction is initially at the concentration of adding maltose in the reactor, 400mmol/L lauric acid acetone soln 250ml, 80g/L lipase, its afterreaction add 20g/L maltose every day, the lauric acid acetone soln of 400mmol/L enters reactor with the 0.1ml/min flow velocity, reaction 10d.The product volume that obtains after output and the separation and purification sees Table 4.

Table 4

Claims (2)

1, a kind of method that the maltose laurate is synthesized in enzyme catalysis continuously in organic phase, it is characterized in that in stirred tank, reacting with continuously feeding and continuous discharging functions, with maltose and lauric acid is raw material, lipase is the synthetic maltose laurate of catalyzer, obtain product maltose laurate through separation and purification, manufacturing condition is:

(1) the maltose initial concentration is 25-200mmol/L in the reactor;

(2) subsequently in the reaction process every day add maltose, addition is 8-20g/L;

(3) the lauric acid concentration in the feeding liquid lauric acid acetone soln is 100-400mmol/L;

(4) consumption of catalyzer lipase NOVE435 is 20-80g/L;

(5) adopt continuous reaction, in reaction process with constant flow pump with in the lauric acid acetone soln suction reactor, the reaction solution of equal volume is drawn out of simultaneously, constant flow pump flow velocity 0.08-0.36mL/min reacted 10-12 days;

(6) with the reaction solution underpressure distillation of collecting, reclaim solvent acetone, obtain solid matter simultaneously;

(7) solid that previous step is obtained adopts normal hexane repeatedly to wash purifying, and normal hexane consumption by volume after the product vacuum-drying that obtains after the washing, is maltose laurate product for 0.1-0.5 times of the reaction solution volume of collection.

2, a kind of method that the maltose laurate is synthesized in enzyme catalysis continuously in organic phase according to claim 1 is characterized in that processing condition are:

(1) the maltose initial concentration is 50mmol/L in the reactor;

(2) subsequently in the reaction process every day add maltose, addition is 16g/L;

(3) the lauric acid concentration in the feeding liquid lauric acid acetone soln is 200mmol/L;

(4) consumption of catalyzer lipase NOVE435 is 40g/L;

(5) adopt continuous reaction, in reaction process with constant flow pump with in the lauric acid acetone soln suction reactor, the reaction solution of equal volume is drawn out of simultaneously, constant flow pump flow velocity 0.15mL/min reacted 10-12 days;

(6) with the reaction solution underpressure distillation of collecting, reclaim solvent acetone, obtain solid matter simultaneously;

(7) solid that previous step is obtained adopts normal hexane washing purifying 3 times, and normal hexane consumption by volume is 0.25 times of the reaction solution volume collected for the first time, and the back is 0.1 times twice, after the product vacuum-drying that obtains after the washing, is maltose laurate product.

Images