CN113749178A - Enzyme treatment method of beta-lactam antibiotic and intermediate bacterial dregs thereof - Google Patents
Enzyme treatment method of beta-lactam antibiotic and intermediate bacterial dregs thereof Download PDFInfo
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- CN113749178A CN113749178A CN202010521442.6A CN202010521442A CN113749178A CN 113749178 A CN113749178 A CN 113749178A CN 202010521442 A CN202010521442 A CN 202010521442A CN 113749178 A CN113749178 A CN 113749178A
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Abstract
The invention belongs to the technical field of antibiotic fungi residue treatment, and particularly relates to a treatment method of beta-lactam antibiotic and intermediate fungi residues thereof. The whole treatment process has mild conditions, low energy consumption and low treatment cost. A large amount of chemical reagents are not added in the treatment process, and the treated dry mushroom dregs have no antibiotic medicine residues, can be used as a protein additive of animal feed, realizes resource recycling, and has good economic benefit, environmental benefit and social benefit.
Description
Technical Field
The invention belongs to the technical field of antibiotic bacteria residue treatment, and particularly relates to a treatment method of beta-lactam antibiotic and intermediate bacteria residue thereof.
Background
Since 1928 Alexada Freimine invented penicillin, the human ability to resist bacterial infection was greatly improved, and penicillin not only saved countless lives, but also led the development of the antibiotic industry. However, with the large-scale use of antibiotics and their abuse, more and more bacteria develop resistance, for example, only 8% of the grape bacteria are resistant to penicillin G when it is first used. By 1962, the number of drug-resistant staphylococcal species increased to 70%. The method for overcoming the drug resistance of bacteria is to develop and use a new semi-synthetic antibiotic, which has the advantages of less side effect, lower toxicity, larger selectivity to pathogenic microorganisms, wide antimicrobial spectrum and the like.
The discovery of 6-aminopenicillanic acid (6-APA), a parent penicillin, has opened a new phase for chemical structural modification of existing antibiotics by chemical methods. The 6-aminopenicillanic acid is a white flaky crystal, is an important intermediate for synthesizing various semi-synthetic penicillins, has wide application range, and is mainly used for synthesizing ampicillin, amoxicillin, phenoxymethylpenicillin, aminopenicillin cloxacillin, mezlocillin, ticarcillin and the like, and other various semi-synthetic penicillins with wider antibacterial spectrum. 7-amino-3-deacetoxy cephalosporanic acid (7-amino des-acetyl cephalosporanic acid, 7-ADCA)) is an important intermediate for synthesizing cephalexin, ceftazidime, cefaclor, cefetamet pivoxil, cefadroxil and other medicaments. 7-aminocephalosporanic acid (7-ACA) is an important intermediate for the production of cephalosporin antibiotics. At present, the parent nucleus of the antibiotics is mainly prepared by deacylation of natural penicillin and cephalosporin C by a chemical method or an enzymatic method. A large amount of toxic reagents are used in the production process of the chemical method, the environment causes certain pollution, and the enzymatic cracking has the advantages of mild condition, high specificity, small environmental pollution, low cost and the like. According to different acylase forms, the method is divided into a solid-phase enzyme method, a hypha suspension method and an immobilized cell method.
Either the fermentative production of penicillins and cephalosporins or the use of enzyme-producing microorganisms to prepare beta-lactam antibiotic intermediates, produce either a bacterial residue or a medicinal residue. China is a large country for producing beta-lactam antibiotics, and over 1 million tons of dry fungus residues are produced every year. Wherein the nontoxic and harmless effective resources account for 99.9 percent of the total mass of the dry fungus dregs and can be further recycled. However, antibiotics with a mass fraction of only 0.1% cannot be decomposed, and recycling of dry mushroom dregs is prohibited, so that the antibiotic dregs cannot be directly used as a raw material of biological feed.
According to investigation, each pharmaceutical enterprise producing beta-lactam antibiotic drugs produces fungi residues in an amount ranging from tens of thousands of tons to hundreds of thousands of tons every year, and the large amount causes great difficulty in fungi residue treatment. For a long time, methods for economically, efficiently and thoroughly treating mushroom dregs have been actively sought. At present, the domestic and foreign treatment methods for the mushroom dregs mainly comprise the following methods: 1) agricultural and animal husbandry utilization: since the mushroom dregs contain a large amount of valuable nutrient elements such as polysaccharide, protein, various amino acids, trace elements and the like, the mushroom dregs are widely used as feed, additives or fertilizers in the early period. The bacterial dregs after high-temperature treatment are used for producing livestock and poultry feed and agricultural fertilizer abroad. However, trace amounts of antibiotics remaining in the mushroom dregs may be enriched in animals and plants, thereby potentially affecting and risking ecosystem, food safety, human health, and the like. In 1/8/2008, antibiotic residues are definitely listed as "dangerous waste" in China, and are prohibited from being directly used as feed and fertilizer; 2) and (3) incineration: the incineration disposal of the bacterial slag can quickly convert the bacterial slag into ash, thereby achieving the purposes of reduction and harmlessness, but the drying and incineration of the bacterial slag need to consume a large amount of heat energy, so the incineration cost is high, the treatment capacity is huge, and the realization difficulty is large; 3) burying: landfill is a mature technology, but the environmental protection problems such as pollution to underground water and the like are prominent, and the simple treatment mode is more and more difficult to continue along with the improvement of attention and requirements of people on the environmental protection problems. No matter which treatment mode is adopted, the burden of treating the medicine dregs of an enterprise is obviously increased, and the resource waste is caused.
The antibiotic residues have high water content and complex components, and contain a large amount of mycoprotein (accounting for more than 40 percent of dry weight), abundant vitamins, crude fat, crude fiber, growth factors, culture medium residues and other nutrient substances. However, trace antibiotics are left in the mushroom dregs, so that the mushroom dregs have a wide antibacterial spectrum, and if the mushroom dregs are not properly treated to enter the environment, the normal growth and multiplication of certain microbial species and groups in the environment can be inhibited, the inherent ecological balance of the environment is damaged, potential harm is caused to the ecological environment, and the mushroom dregs can enter a food chain when serious, so that the health of human beings is harmed.
The standardized disposal of antibiotic fungi residues becomes one of the important factors restricting the development of the pharmaceutical industry at present, and the realization of resource utilization while carrying out harmless treatment on the antibiotic fungi residues becomes the key point of urgent research and development. Wherein the harmless treatment is that the antibiotic residue, harmful and toxic pollution microorganisms, inorganic salt and other substances are reduced below national safety indexes or completely eliminated, so that the antibiotic dregs become substances which are nontoxic and harmless to human bodies and ecological environments; the resource treatment refers to the step of decomposing the mixture after the harmless treatment into further basic materials and intermediates which can meet the production requirements of probiotics, enzyme preparations, other various biotechnological products and biochemical engineering products, and can provide core raw material substances for the mass production of subsequent biological products. For example, the biological feed prepared by taking the antibiotic dregs as the raw materials does not contain antibiotics basically, can effectively improve the quality of livestock and poultry products, can improve the immunocompetence of the livestock and poultry, ensures the livestock and poultry to grow robustly and quickly, increases the product yield, and meets the green, healthy and safe living needs.
Chinese patent application CN01106380.7 discloses a detoxified penicillin fungi residue, a preparation method thereof and application of the detoxified penicillin fungi residue as mycoprotein in feed. The preparation method comprises the following steps: preparation of strains: the used strains are beer yeast and rhodopseudomonas, the beer yeast and the rhodopseudomonas are cultured in a liquid culture medium in a composite way, and the culture conditions are as follows: intermittently aerating at 15-35 ℃ with aeration rate of 1: 0.4-1: 0.8vvm, intermittently stirring at a stirring speed of 100-220 rpm for 1-5 days; secondly, pretreatment: adding acid liquor such as hydrochloric acid or other acid liquor into the wet penicillin fungi residues to adjust the pH value to 2-5, treating for 4-8 hours at the temperature of 60-80 ℃, and neutralizing with alkali liquor such as sodium hydroxide solution or other alkali liquor to adjust the pH value to 6.0-6.5; fermenting in a fermentation tank: putting the processed wet mushroom dregs into a fermentation tank, wherein the charging amount accounts for 60-80% of the fermentation volume, adding a carbon source substance such as glucose or other saccharide substances accounting for 1-10% of the weight of the mushroom dregs, sterilizing at 121 ℃ for 30 minutes, inoculating 0.5-25% of seed liquid when the temperature is reduced to 30 +/-2 ℃, intermittently ventilating at 15-35 ℃, the ventilation amount is 1: 0.4-1: 0.8vvm, intermittently stirring, culturing for 2-7 days at the stirring speed of 100-220 rpm, and putting the fermentation liquid into a storage tank; spray drying: controlling the inlet temperature at 160-190 ℃, and collecting dry powder to obtain the finished product of the detoxified penicillin fungi residue. The invention solves the problems that the penicillin fungi residue is difficult to detoxify and dry thoroughly, the feeding safety is low, the environment is polluted and the like. However, the preparation of the detoxified penicillin fungi residue needs to be carried out in advance, and the preparation method is complicated and is not easy to operate; carbon source substances are also required to be added during fermentation treatment, so that the treatment cost is increased; and the treated product does not detect the residual amount of penicillin, and still has safety risk when being used for feed.
There are also many studies on treating penicillin fungi residues by adopting an anaerobic fermentation mode, but the existing penicillin fungi residue fermentation treatment method generally has the defects of difficult continuous fermentation, incomplete treatment, difficult complete removal of drug residues and the like, and the application of the penicillin fungi residue fermentation treatment method is limited.
Chinese patent application 201410064564.1 discloses a harmless treatment method of fermented antibiotic dregs, which comprises three steps of stirring treatment, steam explosion treatment and drying treatment, wherein antibiotic dregs and plant fiber dregs are fully stirred and uniformly mixed according to the weight ratio of 1: 1; feeding the mixed and stirred pretreatment material into a steam explosion device preheated to more than 150 ℃ to obtain a mixed material after steam explosion through the pressure and temperature of high-pressure steam; and feeding the mixed material subjected to steam explosion into a dryer for drying and crushing. The invention provides a method for innocent treatment of fermented antibiotic residues and subsequent processing of the fermented antibiotic residues serving as raw materials, but the method relates to high-energy-consumption operation such as high temperature, gas explosion and the like, and whether the antibiotic residues are controlled or not is not evaluated.
Chinese patent application CN201010593373 discloses an antibiotic residue resistance solving method, the resistance solving principle of which is as follows: placing the actinomycetes and the like in the medicine residue in an environment which is not suitable for survival, and enabling the EM bacteria protected by the oligosaccharide to continue to live, wherein the EM bacteria can kill the actinomycetes and the like in the medicine residue and consume antibiotics; the functions of the various raw materials used are as follows: in the EM original liquid, EM is Effective Microorganisms, namely Effective Microorganisms consisting of more than 5 kinds of 10 kinds of Microorganisms belonging to 80 kinds, such as photosynthetic bacteria, lactic acid bacteria flora, yeast flora, actinomycete flora, filamentous flora and the like. The method needs complex zymophyte preparation and has long treatment period.
Chinese patent CN201210479546 discloses an antibiotic dregs treatment method, which comprises the following steps: antibiotic medicine residue wet residue is taken as a raw material and is subjected to adsorption degradation, neutralization degradation, fermentation sterilization, deodorization detoxification, drying and utilization. The adsorption and degradation adopts the steps that bentonite and medical stone powder with a certain weight proportion are added into wet antibiotic dregs, and the mixture is uniformly stirred in a containing pool and is kept stand for precipitation, so that part of the residual medicine in the wet antibiotic dregs is degraded; the neutralization degradation means that a proper amount of edible alkaline substances are added into the wet acidic antibiotic dregs to neutralize the dregs so as to enable the dregs to be neutral and further degrade and disappear the residual medicine components; adding a proper amount of edible acidic substances and DDS into the alkaline antibiotic dregs for neutralization to make the dregs neutral and further degrade and eliminate the residual medicinal components; adding yeast yellow wine and saccharified DDG in a certain weight ratio into wet antibiotic dregs, and killing harmful bacteria in the wet dregs in the fermentation process, so that beneficial bacteria are multiplied and residual medicine is further degraded; the deodorization and detoxification adopts the steps that a glucose licorice aqueous medicament with a certain weight proportion is added into wet antibiotic dregs, so that the peculiar smell in the wet antibiotic dregs is removed, and the toxicity of residual medicament in the wet antibiotic dregs is further fully cleared up, thereby ensuring that the wet antibiotic dregs are completely and thoroughly treated so as to be effectively utilized or harmlessly discharged; the drying means that all or part of the processed wet antibiotic dregs are dried and dried in the sun, which is convenient for use and storage. The method adopting physical adsorption and pH value degradation has no specificity, has complex operation process, relates to fermentation and prolongs the treatment period.
Chinese patent 201511021128.7 discloses a treatment process of antibiotic fungi residues, which comprises the following steps: degrading antibiotic bacteria residues to be treated in the presence of an acidic substance under a closed condition at 40-260 ℃, separating degradation products to obtain a solid phase substance flow and a gas phase substance flow, and performing aftertreatment on the gas phase substance flow. Through the treatment, the residual antibiotics in the antibiotic residues can be effectively degraded, meanwhile, the organic nutrient components in the antibiotic residues can be effectively reserved, and good precondition is provided for harmless utilization of the residues. The process can effectively avoid the problem that the mushroom dregs release unpleasant smell in the treatment process and after treatment, and the treated mushroom dregs have no odor and are environment-friendly. The method mainly adopts high temperature treatment, has no specificity, and can not completely retain nutrient substances.
In 2009, 9, the application of penicillinase (such as Liuhui silk-tree) in the harmless treatment of penicillin fungi residues is reported in biological processing, the penicillinase is mixed into the fungi residues according to the ratio of the activity to the cost of the residual penicillin being 3: 1, the residual penicillin can be completely degraded after 1 hour at 37 ℃, the harmless treatment of the fungi residues is realized, and a foundation is laid for the resource utilization of the penicillin fungi residues. The specific operation is as follows: taking 500g of penicillin fungi residue, adding 750mL of purified water, uniformly stirring, standing at room temperature for 15min, adding a proper amount of enzyme with a known concentration, reacting for 1h, centrifuging at 9000rpm/min, filtering supernate with a 0.22-micron microporous membrane, and detecting the titer of residual penicillin in the filtrate by HPLC. The method for treating the antibiotic residues in the mushroom dregs has specificity, but the method determines the antibiotic residues in the liquid after the mushroom dregs are centrifuged, and does not explain the antibiotic residues in the solid.
In order to overcome the above problems, it is necessary to develop a simple and feasible method for specifically eliminating residual antibiotics in herb residue, so as to utilize the nutritive value of herb residue, improve the utilization rate of resources, and improve the environmental, economic and social benefits of the antibiotic industry.
Disclosure of Invention
The invention aims to provide a method for treating beta-lactam antibiotic and intermediate bacterial residues thereof, which has the advantages of single action and simple and easy treatment condition, and adopts the following technical scheme for realizing the purpose of the invention:
a process for treating the bacterial dregs as the intermediate of beta-lactam antibiotic includes adding cracking enzyme, thermal insulating treatment, proportionally mixing beta-lactamase with bacterial dregs, thermal insulating treatment under the optimal action of beta-lactamase, baking and pulverizing.
The invention relates to a method for treating beta-lactam antibiotic and intermediate bacterial residues thereof, which is characterized in that muramidase, cellulase and xylanase are added, the heat preservation condition is 15-40 ℃, the time is 1-24 hours, the ratio of the muramidase to the bacterial residues is 1: 500-1: 10000, and the antibiotic or the intermediate thereof in bacteria can be released through the treatment of the muramidase, so that the next step of the full action of beta-lactamase is facilitated.
The invention relates to a method for treating beta-lactam antibiotics and intermediate mushroom dregs thereof, which comprises the steps of treating and preserving heat for 6-12 hours under the condition of 30-37 ℃ by using a cracking enzyme, wherein the ratio of the cracking enzyme to the mushroom dregs is 1: 1000-1: 5000.
The beta-lactamase used in the method for treating the beta-lactam antibiotic and the intermediate bacterial dregs thereof is penicillinase or cephalosporinase. The final concentration of the penicillinase is 200 units/ml to 5000 units/ml, and the final concentration of the cephalosporinase is 20 units/ml to 500 units/ml.
The beta-lactamase used in the method for treating the beta-lactam antibiotic and the intermediate bacterial dregs thereof is obtained by extracting after the culture induction of enzyme-producing microorganisms or is prepared after the recombinant expression of the gene engineering technology.
The invention relates to a method for treating beta-lactam antibiotic and intermediate bacterial dregs thereof, wherein penicillin dregs and 6-aminopenicillanic acid dregs are treated by penicillinase, and cephalosporin dregs, 7-amino-3-desacetoxy cephalosporanic acid dregs and 7-aminopenicillanic acid dregs are treated by cephalosporinase.
The invention relates to a method for treating beta-lactam antibiotic and intermediate bacterial residue thereof, wherein the optimum action condition of beta-lactamase is pH 6-8, the optimum action temperature is 30-37 ℃, the ionic strength is less than 100mM, and the electric conductivity is less than 10 mS/cm.
The optimal action condition of the optimized beta-lactamase is pH7.0, the optimal action temperature is 35-37 ℃, the ionic strength is less than 50mM, and the electric conductivity is less than 5 mS/cm.
According to the method for treating the beta-lactam antibiotic and the intermediate bacterial sludge thereof, due to the adoption of the action of the specific enzyme, the action of the antibiotic remained in the treated bacterial sludge is specific, the use of the wall-splitting enzyme ensures that the treatment effect is more thorough, and the whole treatment process has mild conditions, low energy consumption and low treatment cost. A large amount of chemical reagents are not added in the treatment process, the treated dry mushroom dregs have no drug residues, and the dry mushroom dregs can be used as a protein additive of animal feed, so that the resource is recycled, and the method has good economic benefit, environmental benefit and social benefit.
Drawings
FIG. 1, RP-HPLC analysis results of penicillin control
FIG. 2 shows the RP-HPLC analysis results of the supernatant after the enzyme treatment of penicillin fungi residues
FIG. 3 shows the RP-HPLC analysis results of the precipitate after the enzyme treatment of penicillin fungi residue
FIG. 4 shows the RP-HPLC analysis results of penicillin fungi residue enzyme treatment, precipitation and cracking of cell wall enzyme treatment
FIG. 5, 6-Effect of APA on E.coli growth;
solid content: adding an enzyme group;
■: the enzyme group was not added.
FIG. 6 shows the results of TLC analysis after enzymatic treatment of mushroom dregs
1: adding no enzyme sample into the mushroom dregs;
2: adding enzyme into the mushroom dregs at 1000u/ml, and processing the mushroom dregs for 1 hour at 37 ℃;
3: adding 10000u/ml enzyme into the mushroom dregs, and processing the mushroom dregs for 1 hour at 50 ℃;
4: 6-APA control sample;
5: 6-APA and penicillin sodium mixed penicillinase are processed for 1 hour at 37 ℃;
6: penicillin sodium control sample.
DETAILED DESCRIPTION OF EMBODIMENT (S) OF INVENTION
The present invention is further illustrated by, but is not limited to, the following examples.
EXAMPLE one penicillinase treatment of penicillin Mushroom dregs
Taking 1KG of penicillin fungi residues, adding 1000mL of water, uniformly stirring, preheating and preserving heat at 37 ℃, adding 2000 ten thousand units/mL of penicillinase to ensure that the final concentration is 1000 units/mL, preserving heat at 37 ℃ for 8 hours, centrifuging, taking supernate, filtering the supernate by a 0.22 mu m microporous filter membrane, and detecting residual penicillin in the filtrate by HPLC, wherein the RP-HPLC analysis method comprises the following steps: c18 column, mobile phase a: 0.1% TFA-H2O, mobile phase B: 0.1% TFA-CAN, gradient 0-5 min, 0% B, 5-15 min, 0-20B, 15-20 min 20% -70% B. The results of the penicillin control degradation are shown in FIG. 1. The analysis result of the bacterial dreg supernatant is shown in figure 2, and penicillin is completely decomposed and can not be detected. And (3) carrying out centrifugal precipitation ultrasonic treatment on the bacterial residues, outputting the power of 100w and 70 percent for 15min, centrifuging after ultrasonic treatment, taking supernate, filtering by using a 0.22-micron microporous filter membrane, and detecting the residual penicillin in the filtrate by using HPLC (high performance liquid chromatography), wherein the result is shown in figure 3, and the penicillin residue in the bacterial residues can be detected.
Example treatment of penicillin fungi residue with muramidase and penicillinase
Taking 1KG of penicillin fungi residues, adding 1000mL of water, uniformly stirring, preheating at 37 ℃, preserving heat, adding 2g of muramidase, wherein 1g of lysozyme, 0.5g of cellulase and 0.5g of xylanase, uniformly mixing, preserving heat at 37 ℃ for 8 hours, adding 2000 kilomega units/mL of penicillinase to ensure that the final concentration is 1000 units/mL, continuously preserving heat at 37 ℃ for 8 hours, centrifuging, taking fungi residues for centrifugation and precipitation, carrying out ultrasonic treatment, outputting power of 100w, 70% and 15min, centrifuging after ultrasonic treatment, taking supernatant, carrying out HPLC (high performance liquid chromatography) detection on residual penicillin in filtrate after filtering through a 0.22 mu m microporous membrane, and detecting penicillin residues with the result shown in figure 4.
EXAMPLES triple lywallzyme and penicillinase treatment of 6-APA fungal dregs
Taking 1KG of 6-APA bacterial residues, adding 1000mL of water, uniformly stirring, preheating and preserving heat at 37 ℃, adding 1g of muramidase, wherein 0.4g of lysozyme, 0.3g of cellulase and 0.3g of xylanase, uniformly mixing, preserving heat at 37 ℃ for 12 hours, adding 2000 ten thousand units/mL of penicillinase to enable the final concentration to be 500 units/mL, continuously preserving heat at 37 ℃ for 10 hours, centrifuging, taking bacterial residues, performing ultrasonic treatment on centrifugal precipitation, outputting 100w and 70 percent of power, performing centrifugal treatment for 15 minutes after ultrasonic treatment, centrifuging to obtain supernatant, filtering through a microporous filter membrane of 0.22 mu m, and detecting residual 6-APA in filtrate by HPLC (high performance liquid chromatography), wherein the result shows that no penicillin residue can be detected.
Examples treatment of cephalosporin sludge with Fomitosin and cephalosporin enzymes
Taking 500g of cephalosporin dregs, adding 750mL of water, uniformly stirring, preheating at 37 ℃, carrying out heat preservation, adding 1g of lyase, wherein 0.35g of lysozyme, 0.35g of cellulase and 0.3g of xylanase, uniformly mixing, carrying out heat preservation treatment at 37 ℃ for 10 hours, adding 200 ten thousand units/mL of cephalosporin enzyme to ensure that the final concentration is 800 units/mL, continuously carrying out heat preservation at 37 ℃ for 12 hours, centrifuging, taking the fungus dregs for ultrasonic treatment, outputting 100w, 70 percent and 15 minutes, centrifuging after ultrasonic treatment, taking supernatant, filtering by a 0.22 mu m microporous filter membrane, and detecting residual cephalosporin in filtrate by HPLC (high performance liquid chromatography), wherein the result is that no cephalosporin residue is detected.
Example treatment of 7-ACA fungal dregs with Pentacrolimus and cephalosporin enzymes
Taking 500g of 7-ACA bacterial residues, adding 500mL of water, uniformly stirring, preheating and preserving heat at 37 ℃, adding 0.5g of muramidase, wherein 0.15g of lysozyme, 0.2 g of cellulase and 0.15g of xylanase, uniformly mixing, preserving heat at 37 ℃ for 10 hours, adding 200 ten thousand units/mL of cephalosporin enzyme to ensure that the final concentration is 1000 units/mL, continuously preserving heat at 37 ℃ for 8 hours, centrifuging, precipitating and ultrasonically treating the bacterial residues, outputting 100w, 70 percent and 15 minutes, centrifuging after ultrasonic treatment, taking supernatant, filtering by a 0.22 mu m microporous filter membrane, and detecting residual 7-ACA in filtrate by HPLC (high performance liquid chromatography), wherein 7-ACA residue cannot be detected.
Example six 6-Effect of APA on the growth of E.coli and the action of penicillinase
6APA solutions with different concentrations are prepared by using sterile LB culture medium, a single colony is picked from an escherichia coli plate and placed in 5ml of sterile LB culture medium to be used as a strain by shaking culture at 37 ℃ and 250rpm, 40 mu l of the single colony is taken under the aseptic condition to inoculate 5ml of LB culture medium containing 6APA, the other group is added with penicillinase to make the final concentration be 10000U/ml, and the solution is shaken at 37 ℃ and 250rpm for 4 hours, and the absorption value of OD600nm is measured, and the result is shown in figure 5. The experimental results show that the 6-APA standard sample has an inhibition effect on the growth of Escherichia coli in LB culture medium when the concentration is more than 50 mu g/ml, but the inhibition effect is not complete. When penicillinase is added, there is no inhibition of E.coli growth due to enzymatic decomposition of 6-APA.
EXAMPLE seven 6-titration of enzymatic hydrolysate after enzymatic treatment of APA Mushroom dregs
Taking the bacterial residue with the pH value of 7.0, centrifuging (4 ℃, 8000rpm, 10min), filtering, adding penicillinase to enable the final concentration to be 10000u/ml, oscillating at 37 ℃, and keeping the temperature at 250rpm for 1 hour, taking 3ml, adding 10ml of iodometric titration solution according to a method of appendix 98 of second part of Chinese pharmacopoeia, diluting to 25ml by using acetic acid buffer solution, reacting for 15min in a dark place, titrating by using sodium thiosulfate titration solution, and taking no enzyme group as a control. The results are shown in Table I.
TABLE I titration results of the penicillinase action product
Penicillinase hydrolyses 6-APA to yield 6-aminopenicillanic acid, which may be reacted with iodine, and excess iodine may be titrated with sodium thiosulfate. The experimental results show that 6-APA in the dregs can be hydrolyzed by penicillinase to generate 6-aminopenicillanic acid, and the latter reacts with iodine to consume a part of iodine, so that the consumed sodium thiosulfate is less than that of a control.
Example Ochrose-residue enzymatic treatment TLC analysis
The bacterial residue with pH7.0 was adjusted to a final concentration of 500u/ml, 1000u/ml, 10000u/ml by adding penicillinase at 37 ℃ and 50 ℃ respectively, and the incubation time was 2 hours, 4 hours, and 6 hours. 10ul of thin-layer chromatography sample application is carried out, the thin-layer chromatography condition for analyzing penicillin potassium is referred, and the developing agent is: shaking mixed solution of butyl acetate, glacial acetic acid, 5% sodium dihydrogen phosphate solution of 0.1% EDTA-2Na and n-butanol at ratio of 60: 20: 10: 5, standing for layering, and collecting upper layer solution. Color developing agent: 10% ninhydrin in methanol. The control solution was 2mg/mL of 6-APA, 10mg/mL of penicillin sodium, and the results are shown in FIG. 6. The results show that the control group and the enzyme-treated sample are not different because the fermentation component is complex and the thin layer chromatography cannot separate the 6-APA. However, the 6-APA standard sample and the penicillin sodium completely disappear after the enzyme action, which indicates that the penicillinase can effectively decompose the 6-APA and the penicillin sodium.
Claims (8)
1. A method for treating beta-lactam antibiotic and its intermediate bacterial dregs comprises mixing beta-lactamase with bacterial dregs uniformly according to a proportion, preserving heat and drying, and is characterized in that: adding a wall-cracking enzyme for heat preservation treatment, and carrying out heat preservation treatment under the optimal action condition of beta-lactamase.
2. The method for treating the beta-lactam antibiotic and the intermediate mushroom dregs thereof according to claim 1, wherein the method comprises the following steps: the added muramidase is lysozyme, cellulase and xylanase, the heat preservation condition is 15-40 ℃, the time is 1-24 hours, and the ratio of the muramidase to the fungus residues is 1: 500-1: 10000.
3. The method for treating the beta-lactam antibiotic and the intermediate mushroom dregs thereof according to claim 1, wherein the method comprises the following steps: the heat preservation condition of the wall cracking enzyme treatment is 30-37 ℃, the time is 6-12 hours, and the ratio of the wall cracking enzyme to the fungus residues is 1: 1000-1: 5000.
4. The method for treating the beta-lactam antibiotic and the intermediate mushroom dregs thereof according to claim 1, wherein the method comprises the following steps: the beta-lactamase is penicillinase or cephalosporinase.
5. The method for treating the beta-lactam antibiotic and the intermediate mushroom dregs thereof according to claim 1, wherein the method comprises the following steps: the beta-lactamase is obtained by extracting after the culture induction of enzyme-producing microorganisms or is prepared after the recombination expression by a genetic engineering technology.
6. The method for treating the beta-lactam antibiotic and the intermediate mushroom dregs thereof according to claim 1, wherein the method comprises the following steps: the penicillin dregs and 6-aminopenicillanic acid dregs are treated by penicillinase, and the cephalosporin dregs, 7-amino-3-desacetoxycephalosporanic acid dregs and 7-aminopenicillanic acid dregs are treated by cephalosporinase.
7. The method for treating the beta-lactam antibiotic and the intermediate mushroom dregs thereof according to claim 1, wherein the method comprises the following steps: the optimum action condition of the beta-lactamase is pH 6-8, the optimum action temperature is 30-37 ℃, the ionic strength is less than 100mM, and the electric conductance is less than 10 mS/cm.
8. The method for treating the beta-lactam antibiotic and the intermediate mushroom dregs thereof according to claim 1, wherein the method comprises the following steps: the optimum action condition of the beta-lactamase is pH7.0, the optimum action temperature is 35-37 ℃, the ionic strength is less than 50mM, and the electric conductance is less than 5 mS/cm.
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Citations (10)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101624605A (en) * | 2008-07-08 | 2010-01-13 | 武汉烁森生态科技有限公司 | Biological method for processing spiramycin bacteria residue |
| CN101979521A (en) * | 2010-09-26 | 2011-02-23 | 吴鹏 | Complex enzyme preparation special for antibiotic dregs |
| CN103865815A (en) * | 2014-02-25 | 2014-06-18 | 北京观澜科技有限公司 | Method for preparing biological humic acid bacteria agent from biological organic enzymolysis-coupled conversion antibiotic dregs |
| CN103937663A (en) * | 2014-05-08 | 2014-07-23 | 杭州北望生物技术有限公司 | Sterile culture medium plate for environment monitoring of antibiotic production workshop and preparation method of sterile culture medium plate |
| CN104328141A (en) * | 2014-10-30 | 2015-02-04 | 宁夏乙征生物工程有限公司 | Method for treating antibiotic residues by enzymic method |
| CN106267676A (en) * | 2016-09-26 | 2017-01-04 | 齐鲁工业大学 | A kind of antibiotic bacterium dregs xylanase enzyme solution |
| CN108277161A (en) * | 2017-01-05 | 2018-07-13 | 天方药业有限公司 | Waste thallus processing method in a kind of production of spiramvcin and waste thallus circulation utilization method |
| CN110305859A (en) * | 2019-07-31 | 2019-10-08 | 陕西麦可罗生物科技有限公司 | A method of biological nitrogen is produced using kasugarnycin and polyoxin bacteria residue |
| CN114230380A (en) * | 2021-12-07 | 2022-03-25 | 山东睿智医药科技有限公司 | Utilization method of 7-aminocephalosporanic acid production waste |
| CN116083407A (en) * | 2022-12-02 | 2023-05-09 | 杭州俊丰生物工程有限公司 | Thermostable cephalosporin enzyme composition for plate medium addition |
-
2020
- 2020-06-02 CN CN202010521442.6A patent/CN113749178A/en active Pending
Patent Citations (10)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101624605A (en) * | 2008-07-08 | 2010-01-13 | 武汉烁森生态科技有限公司 | Biological method for processing spiramycin bacteria residue |
| CN101979521A (en) * | 2010-09-26 | 2011-02-23 | 吴鹏 | Complex enzyme preparation special for antibiotic dregs |
| CN103865815A (en) * | 2014-02-25 | 2014-06-18 | 北京观澜科技有限公司 | Method for preparing biological humic acid bacteria agent from biological organic enzymolysis-coupled conversion antibiotic dregs |
| CN103937663A (en) * | 2014-05-08 | 2014-07-23 | 杭州北望生物技术有限公司 | Sterile culture medium plate for environment monitoring of antibiotic production workshop and preparation method of sterile culture medium plate |
| CN104328141A (en) * | 2014-10-30 | 2015-02-04 | 宁夏乙征生物工程有限公司 | Method for treating antibiotic residues by enzymic method |
| CN106267676A (en) * | 2016-09-26 | 2017-01-04 | 齐鲁工业大学 | A kind of antibiotic bacterium dregs xylanase enzyme solution |
| CN108277161A (en) * | 2017-01-05 | 2018-07-13 | 天方药业有限公司 | Waste thallus processing method in a kind of production of spiramvcin and waste thallus circulation utilization method |
| CN110305859A (en) * | 2019-07-31 | 2019-10-08 | 陕西麦可罗生物科技有限公司 | A method of biological nitrogen is produced using kasugarnycin and polyoxin bacteria residue |
| CN114230380A (en) * | 2021-12-07 | 2022-03-25 | 山东睿智医药科技有限公司 | Utilization method of 7-aminocephalosporanic acid production waste |
| CN116083407A (en) * | 2022-12-02 | 2023-05-09 | 杭州俊丰生物工程有限公司 | Thermostable cephalosporin enzyme composition for plate medium addition |
Non-Patent Citations (1)
| Title |
|---|
| 刘慧娟等: "青霉素酶在青霉素菌渣无害化处理中的应用", 《生物加工过程》, vol. 7, no. 5, pages 54 - 57 * |
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