Cyclosporin A fermentation residue(CFR)is a type of organic waste generated during the production of cyclosporin A,which are abundant in nutrients including organic matter,phosphorus,nitrogen and potassium.Inappropria...Cyclosporin A fermentation residue(CFR)is a type of organic waste generated during the production of cyclosporin A,which are abundant in nutrients including organic matter,phosphorus,nitrogen and potassium.Inappropriate handling of CFR not only waste valuable bioresources,but may also lead to the cyclosporin A and associated resistance genes into the natural environment,posing a significant threat to ecological system and human health.Land application was an effective way to resource recovery of CFR after aerobic composting(CAC).This study investigated the impact of CAC on soil fertility and environmental safety.The results indicated that CAC could improve soil nutrient contents and enhance enzyme activities.CAC altered the diversity and community composition of soil bacteria,resulting in an increase in the abundance of relevant bacteria beneficial for organic matter decomposition and cyclosporin A degradation.The introduced cyclosporin A(71.69μg/kg)completely degraded within 20 days due to soil biodegradation.The significantly increased abundance of intIl,mdr3,pgp,TSR and pmra in the soil cultivation early stage were restored to the soil background level within 90 days,indicating a reduced risk of antimicrobial resistance.The results demonstrated that reasonable land application of CAC could improve soil fertility without antimicrobial resistance risk,which is helpful for evaluating the resource utilization value and environmental risks of antibiotic fermentation residue after aerobic composting.展开更多
Erythromycin fermentation residue(EFR)represents a typical hazardous waste produced by the microbial pharmaceutical industry.Although electrolysis is promising for EFR disposal,its microbial threats remain unclear.Her...Erythromycin fermentation residue(EFR)represents a typical hazardous waste produced by the microbial pharmaceutical industry.Although electrolysis is promising for EFR disposal,its microbial threats remain unclear.Herein,metagenomics was coupled with the random forest technique to decipher the antibiotic resistance patterns of electrochemically treated EFR.Results showed that 95.75%of erythromycin could be removed in 2 hr.Electrolysis temporarily influenced EFRmicrobiota,where the relative abundances of Proteobacteria and Actinobacteria increased,while those of Fusobacteria,Firmicutes,and Bacteroidetes decreased.A total of 505 antibiotic resistance gene(ARG)subtypes encoding resistance to 21 antibiotic types and 150 mobile genetic elements(MGEs),mainly including plasmid(72)and transposase(52)were assembled in EFR.Significant linear regression models were identified among microbial richness,ARG subtypes,and MGE numbers(r^(2)=0.50-0.81,p<0.001).Physicochemical factors of EFR(Total nitrogen,total organic carbon,protein,and humus)regulated ARG and MGE assembly(%IncMSE value=5.14-14.85).The core ARG,MGE,and microbe sets(93.08%-99.85%)successfully explained 89.71%-92.92%of total ARG and MGE abundances.Specifically,gene aph(3 )-I,transposase tnpA,and Mycolicibacterium were the primary drivers of the resistance dissemination system.This study also proposes efficient resistance mitigation measures,and provides recommendations for future management of antibiotic fermentation residue.展开更多
Aim A novel method has been developed for evaluation of the levels of total residual protein in antibiotics produced by fermentation using gel filtration chromatography (GFC) combined with Bradford assay based on dete...Aim A novel method has been developed for evaluation of the levels of total residual protein in antibiotics produced by fermentation using gel filtration chromatography (GFC) combined with Bradford assay based on determination of residual protein in lincomycin hydrochloride. Methods The chromatographic conditions were SuperdexTM peptide column, 0.01 mol*L-1 phosphate buffer solution as mobile phase, and flow rate of 1 mL·min-1. Five hundred microliters of lincomycin hydrochloride solution (3 g of lincomycin hydrochloride dissolved in 10 mL of mobile phase) was injected into the chromatograph and the eluted solution was collected between 6 min and 14.5 min (protein eluted from column within this period), and the residual content of total protein in the eluted solution was assayed using Bradford assay method. Results The average recovery was more than 90% for bovine serum albumin, the calibration equation for the range of 0-12 μg·mL-1 of protein was y=-0.002 4x2+0.064 2x+0.002 9, r2=0.999 9, RSD=0.1%-0.9%, and the LOD and LOQ were 3 and 10 ng·mL-1 of protein, respectively. Conclusion The novel method for determining the residual protein in ferment antibio-tics is simple, rapid, and precise.展开更多
Monensin,a polyether ionophore antibiotic that is produced by Streptomyces cinnamonensis through fermentation,is extensively utilized in both agricultural and pharmaceutical sectors.This study focused on identifying s...Monensin,a polyether ionophore antibiotic that is produced by Streptomyces cinnamonensis through fermentation,is extensively utilized in both agricultural and pharmaceutical sectors.This study focused on identifying some specific genes and critical metabolic pathways related to the monensin biosynthesis in S.cinnamonensis for efficient monensin production with a genome analysis.Results show that genes of the strain were significantly enriched in the monensin synthetic pathway,including primary metabolic(central carbon and fatty acids)processes,energy metabolism,and secondary metabolite biosynthesis,which was largely potential in the supply of sufficient building precursors and energy.The annotated specific genes were predominantly located in metabolic pathways and secondary metabolites biosynthesis,accounting for 90.63%and 39.06%,respectively.Among them,specific genes,fadD,fadE,fadB,and fadA in the fatty acid degradation pathway were apparently the most prominent.With single overexpression,these genes resulted in the strain increasing monensin titer by 14%,11%,22%,and 10%,respectively.Further,with the tandem overexpression,an engineered strain M5 was successfully constructed.The strain was capable of producing 18.88 g/L of monensin at 288 h at shake-flask level and 37.31 g/L via fed-batch in a 50-L bioreactor,which is 1.3 folds and 1.2 folds,respectively,that of the starting strain.To our knowledge,this represents the highest level reported to date,which is of a big industrial promise.This study provides a genetic foundation for elucidating the monensin synthesis mechanism and paves the way for metabolic engineering modifications and industrial production.展开更多
基金supported by the National Natural Science Foundation of China(No.52170138)the National Key Research and Development Program of China(No.2023YFD1900305)+3 种基金Qingdao Agricultural University Doctoral Start-Up Fund(No.6631122003)the Project of Talent IntroductionEducation Program of Youth Innovation Teams in Universities of Shandong Province(2021–05)Shandong Provincial College Youth Innovation Team Program(No.2023KJ169).
文摘Cyclosporin A fermentation residue(CFR)is a type of organic waste generated during the production of cyclosporin A,which are abundant in nutrients including organic matter,phosphorus,nitrogen and potassium.Inappropriate handling of CFR not only waste valuable bioresources,but may also lead to the cyclosporin A and associated resistance genes into the natural environment,posing a significant threat to ecological system and human health.Land application was an effective way to resource recovery of CFR after aerobic composting(CAC).This study investigated the impact of CAC on soil fertility and environmental safety.The results indicated that CAC could improve soil nutrient contents and enhance enzyme activities.CAC altered the diversity and community composition of soil bacteria,resulting in an increase in the abundance of relevant bacteria beneficial for organic matter decomposition and cyclosporin A degradation.The introduced cyclosporin A(71.69μg/kg)completely degraded within 20 days due to soil biodegradation.The significantly increased abundance of intIl,mdr3,pgp,TSR and pmra in the soil cultivation early stage were restored to the soil background level within 90 days,indicating a reduced risk of antimicrobial resistance.The results demonstrated that reasonable land application of CAC could improve soil fertility without antimicrobial resistance risk,which is helpful for evaluating the resource utilization value and environmental risks of antibiotic fermentation residue after aerobic composting.
基金supported by the Special Project of Basic Scientific Research Business of Central Public Welfare Scientific Research Institutes (No.2019YSKY-027).
文摘Erythromycin fermentation residue(EFR)represents a typical hazardous waste produced by the microbial pharmaceutical industry.Although electrolysis is promising for EFR disposal,its microbial threats remain unclear.Herein,metagenomics was coupled with the random forest technique to decipher the antibiotic resistance patterns of electrochemically treated EFR.Results showed that 95.75%of erythromycin could be removed in 2 hr.Electrolysis temporarily influenced EFRmicrobiota,where the relative abundances of Proteobacteria and Actinobacteria increased,while those of Fusobacteria,Firmicutes,and Bacteroidetes decreased.A total of 505 antibiotic resistance gene(ARG)subtypes encoding resistance to 21 antibiotic types and 150 mobile genetic elements(MGEs),mainly including plasmid(72)and transposase(52)were assembled in EFR.Significant linear regression models were identified among microbial richness,ARG subtypes,and MGE numbers(r^(2)=0.50-0.81,p<0.001).Physicochemical factors of EFR(Total nitrogen,total organic carbon,protein,and humus)regulated ARG and MGE assembly(%IncMSE value=5.14-14.85).The core ARG,MGE,and microbe sets(93.08%-99.85%)successfully explained 89.71%-92.92%of total ARG and MGE abundances.Specifically,gene aph(3 )-I,transposase tnpA,and Mycolicibacterium were the primary drivers of the resistance dissemination system.This study also proposes efficient resistance mitigation measures,and provides recommendations for future management of antibiotic fermentation residue.
文摘Aim A novel method has been developed for evaluation of the levels of total residual protein in antibiotics produced by fermentation using gel filtration chromatography (GFC) combined with Bradford assay based on determination of residual protein in lincomycin hydrochloride. Methods The chromatographic conditions were SuperdexTM peptide column, 0.01 mol*L-1 phosphate buffer solution as mobile phase, and flow rate of 1 mL·min-1. Five hundred microliters of lincomycin hydrochloride solution (3 g of lincomycin hydrochloride dissolved in 10 mL of mobile phase) was injected into the chromatograph and the eluted solution was collected between 6 min and 14.5 min (protein eluted from column within this period), and the residual content of total protein in the eluted solution was assayed using Bradford assay method. Results The average recovery was more than 90% for bovine serum albumin, the calibration equation for the range of 0-12 μg·mL-1 of protein was y=-0.002 4x2+0.064 2x+0.002 9, r2=0.999 9, RSD=0.1%-0.9%, and the LOD and LOQ were 3 and 10 ng·mL-1 of protein, respectively. Conclusion The novel method for determining the residual protein in ferment antibio-tics is simple, rapid, and precise.
基金funded by the National Natural Science Foundation of China(21776114)the National First-class Discipline Program of Light Industry Technology and Engineering(QGJC20230203)+1 种基金the 111 Project(111-2-06)the Postgraduate Research and Practice Innovation Program of Jiangsu Province(KYCX22-2377).
文摘Monensin,a polyether ionophore antibiotic that is produced by Streptomyces cinnamonensis through fermentation,is extensively utilized in both agricultural and pharmaceutical sectors.This study focused on identifying some specific genes and critical metabolic pathways related to the monensin biosynthesis in S.cinnamonensis for efficient monensin production with a genome analysis.Results show that genes of the strain were significantly enriched in the monensin synthetic pathway,including primary metabolic(central carbon and fatty acids)processes,energy metabolism,and secondary metabolite biosynthesis,which was largely potential in the supply of sufficient building precursors and energy.The annotated specific genes were predominantly located in metabolic pathways and secondary metabolites biosynthesis,accounting for 90.63%and 39.06%,respectively.Among them,specific genes,fadD,fadE,fadB,and fadA in the fatty acid degradation pathway were apparently the most prominent.With single overexpression,these genes resulted in the strain increasing monensin titer by 14%,11%,22%,and 10%,respectively.Further,with the tandem overexpression,an engineered strain M5 was successfully constructed.The strain was capable of producing 18.88 g/L of monensin at 288 h at shake-flask level and 37.31 g/L via fed-batch in a 50-L bioreactor,which is 1.3 folds and 1.2 folds,respectively,that of the starting strain.To our knowledge,this represents the highest level reported to date,which is of a big industrial promise.This study provides a genetic foundation for elucidating the monensin synthesis mechanism and paves the way for metabolic engineering modifications and industrial production.
