聚乙烯醇降解细菌筛选及其降解特性
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摘要
从聚乙烯醇泡棉的堆肥降解过程中筛选出可以有效降解聚乙烯醇的菌株,并研究其对聚乙烯醇的降解效果和降解特性。将聚乙烯醇泡棉经过3年的堆肥降解,利用傅里叶转换红外光谱(Fourier transform infrared spectroscopy,FTIR)检测泡棉的分子结构变化;采用Finley法从降解后的聚乙烯醇泡棉中筛选降解效果较好的菌株,通过菌落形态和细胞染色以及16S rRNA基因序列比对进行鉴定;通过紫外分光光度计检测菌株在降解聚乙烯醇材料过程中的菌体浓度变化和聚乙烯醇降解程度,并将筛选菌株对两种醇解度的聚乙烯醇降解效果进行对比。结果表明,经过3年的堆肥降解,聚乙烯醇泡棉分子结构中出现了新的羧基和羟基;筛选得到了四株菌,即Bacillus sp.DG01、Bacillus sp.DG02、Paenibacillus sp.DG03、Paenibacillus sp.DG04。这4株筛选菌株对3.0g/L PVA1788的降解率分别为67.27%、74.99%、56.74%和59.70%,对3.0 g/L PVA1799的降解率分别为58.27%、54.47%、43.32%和46.59%。PVA泡棉在堆肥降解过程中发生了明显的基团变化,4株菌的生长与聚乙烯醇的降解过程呈耦联关系,且4株菌株整体上对较低醇解度的PVA降解效果更好。
The aim of this work is to isolate and identify the strains which can effectively degrade polyvinyl alcohol(PVA)from the composting degradation process of PVA foams and further to study the degradation effects and characteristics of these strains on PVA. First,the PVA foam was degraded after 3 years composting,and we used Fourier Transform Infrared Spectroscopy(FTIR)to detect the molecular structure change of the foam. Then we used Finley method to screen the strains with better degradability from the degraded foams. Further we combined morphological characteristics,gram staining experiment and 16 s rRNA gene sequence alignment to identify the strains. Finally we employed UV ultraviolet spectrophotometer to measure the degradation rates of PVA by various strains and the OD600 of cells in shake flask,and we also compared the degradability of PVA by the screened strains at 2 alcoholysis degrees. The results showed new carboxyl groups and hydroxyl groups appeared in the molecular structure of the PVA foam after 3 years composting degradation;4 strains were screened and identified,including Bacillus sp. DG01,Bacillus sp. DG02,Paenibacillus sp. DG03 and Paenibacillus sp. DG04. The degradation rates by these 4 isolated strains at 3.0 g/L PVA1788 were 67.27%,74.99%,56.74% and 59.70%,respectively,and the degradation rates at 3.0 g/L PVA1799 were 58.27%,54.47%,43.32%,and 46.59%,respectively. The PVA foams presented obvious group changes after the composting;the growth of 4 strains was coupled with the degradation of PVA,and the low alcoholysis degree of PVA was more beneficial to the biodegradation.
The aim of this work is to isolate and identify the strains which can effectively degrade polyvinyl alcohol(PVA)from the composting degradation process of PVA foams and further to study the degradation effects and characteristics of these strains on PVA. First,the PVA foam was degraded after 3 years composting,and we used Fourier Transform Infrared Spectroscopy(FTIR)to detect the molecular structure change of the foam. Then we used Finley method to screen the strains with better degradability from the degraded foams. Further we combined morphological characteristics,gram staining experiment and 16 s rRNA gene sequence alignment to identify the strains. Finally we employed UV ultraviolet spectrophotometer to measure the degradation rates of PVA by various strains and the OD600 of cells in shake flask,and we also compared the degradability of PVA by the screened strains at 2 alcoholysis degrees. The results showed new carboxyl groups and hydroxyl groups appeared in the molecular structure of the PVA foam after 3 years composting degradation;4 strains were screened and identified,including Bacillus sp. DG01,Bacillus sp. DG02,Paenibacillus sp. DG03 and Paenibacillus sp. DG04. The degradation rates by these 4 isolated strains at 3.0 g/L PVA1788 were 67.27%,74.99%,56.74% and 59.70%,respectively,and the degradation rates at 3.0 g/L PVA1799 were 58.27%,54.47%,43.32%,and 46.59%,respectively. The PVA foams presented obvious group changes after the composting;the growth of 4 strains was coupled with the degradation of PVA,and the low alcoholysis degree of PVA was more beneficial to the biodegradation.
引文
[1]雷德松.聚乙烯醇(PVA)生物降解性研究的最新进展[J].中国化工贸易, 2015(15):118-118.
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[4]Matsumura S, Tomizawa N, Toki A, et al. Novel poly(vinyl alcohol)-degrading enzyme and the degradation mechanism[J].Macromolecules, 1999, 32(23):7753-7761.
[5]Liu YK, Zhou YH, Chen SL, et al. Identification and degradation character of a polyvinyl alcohol degradable bacteria HK1[J].Journal of Anhui Agricultural Sciences, 2017, 45(22):5-8, 19.
[6]Maiti S, Ray D, Mitra D, et al. Isolation and characterisation of starch/polyvinyl alcohol degrading fungi from aerobic compost environment[J]. International Biodeterioration&Biodegradation,2013, 82(8):9-12.
[7]Corti A, Solaro R, et al. Biodegradation of poly(vinyl alcohol)in selected mixed microbial culture and relevant culture filtrate[J].Polymer Degradation&Stability, 2002, 75(3):447-458.
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[9]Karamanlioglu M, Houlden A, Robson GD. Isolation and characterisation of fungal communities associated with degradation and growth on the surface of poly(lactic)acid(PLA)in soil and compost[J]. International Biodeterioration&Biodegradation,2014, 95(3):301-310.
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[11]段梦洁.聚乙烯醇材料降解微生物的菌群结构分析及其作用菌株分离[D].广州:华南理工大学, 2018.
[12]Finley JH. Spectrophotometric determination of polyvinyl alcohol in paper coatings[J]. Analytical Chemistry, 1961, 33(13):1925-1927.
[13]段绪果,周素雅,沈镇炎,等.一株产生淀粉酶杆菌Bacillus sp. GEL-09的筛选、鉴定及发酵条件优化[J].微生物学通报.2018, 45(6):1180-1189.
[14]Ullah M, Weng CH, Li H, et al. Degradation of polyvinyl alcohol by a novel bacterial strain Stenotrophomonas sp. SA21[J].Environmental Technology, 2017, 39(16):2056-1061.
[15]董丽娟.聚乙烯醇生物降解性能的研究[D].南京:南京理工大学, 2005.
[16]Lopez BLO, Mejia AIG, Sierra LG. Biodegradability of poly(vinyl alcohol). Polymer Engineering&Science[J]. 2010, 39(8):1346-1352.
[17]Choi K, Park C, Kim S, et al. Polyvinyl alcohol degradation by microbacterium barkeri KCCM 10507 and Paenibacillus amylolyticus KCCM 10508 in dyeing wastewater[J]. Journal of Microbiology&Biotechnology, 2004, 14(5):1009-1013.
[18]Mori T, Sakimoto M, Kagi TT. Isolation and characterization of a strain of bacillus megaterium that degrades poly(vinyl alcohol)[J]. Bioscience Biotechnology&Biochemistry, 1996,60(2):330-332.
[19]Chen J, Zhang Y, Du GC, et al. Biodegradation of polyvinyl alcohol by a mixed microbial culture[J]. Enzymeµbial technology, 2007, 40(7):1686-1691.
[20]赵一凡.聚乙烯醇降解菌的筛选及其关键酶的应用研究[D].无锡:江南大学, 2017.
[21]张颖.微生物降解聚乙烯醇的条件及降解过程机制研究[D].无锡:江南大学, 2006.
[22]厉成宣,范雪荣,王强,等.退浆废水中PVA对环境的影响及其降解性能[J].印染助剂, 2007, 24(6):7-10.
[2]Suzuki T, Ichihara Y, Yamada M, et al. Some characteristics of 0-3which utilizes polyvinyl alcohol[J]. Journal of the Agricultural Chemical Society of Japan, 2014, 37(4):747-756.
[3]Shimao M, Onishi S, Kato N, et al. Pyrroloquinoline quinonedependent cytochrome reduction in polyvinyl alcohol-degrading Pseudomonas sp. Strain VM15C[J]. Applied&Environmental Microbiology, 1989, 55(2):275-278.
[4]Matsumura S, Tomizawa N, Toki A, et al. Novel poly(vinyl alcohol)-degrading enzyme and the degradation mechanism[J].Macromolecules, 1999, 32(23):7753-7761.
[5]Liu YK, Zhou YH, Chen SL, et al. Identification and degradation character of a polyvinyl alcohol degradable bacteria HK1[J].Journal of Anhui Agricultural Sciences, 2017, 45(22):5-8, 19.
[6]Maiti S, Ray D, Mitra D, et al. Isolation and characterisation of starch/polyvinyl alcohol degrading fungi from aerobic compost environment[J]. International Biodeterioration&Biodegradation,2013, 82(8):9-12.
[7]Corti A, Solaro R, et al. Biodegradation of poly(vinyl alcohol)in selected mixed microbial culture and relevant culture filtrate[J].Polymer Degradation&Stability, 2002, 75(3):447-458.
[8]罗洁,郝喜海,李贞,等.类产碱假单胞菌对不同型号PVA的降解特性研究[J].包装学报, 2013(4):34-38.
[9]Karamanlioglu M, Houlden A, Robson GD. Isolation and characterisation of fungal communities associated with degradation and growth on the surface of poly(lactic)acid(PLA)in soil and compost[J]. International Biodeterioration&Biodegradation,2014, 95(3):301-310.
[10]郝喜海,衣潇鹏,李丁,等.聚乙烯醇薄膜降解菌的筛选及助剂的影响研究[J].广州化工, 2015, 43(1):3-6.
[11]段梦洁.聚乙烯醇材料降解微生物的菌群结构分析及其作用菌株分离[D].广州:华南理工大学, 2018.
[12]Finley JH. Spectrophotometric determination of polyvinyl alcohol in paper coatings[J]. Analytical Chemistry, 1961, 33(13):1925-1927.
[13]段绪果,周素雅,沈镇炎,等.一株产生淀粉酶杆菌Bacillus sp. GEL-09的筛选、鉴定及发酵条件优化[J].微生物学通报.2018, 45(6):1180-1189.
[14]Ullah M, Weng CH, Li H, et al. Degradation of polyvinyl alcohol by a novel bacterial strain Stenotrophomonas sp. SA21[J].Environmental Technology, 2017, 39(16):2056-1061.
[15]董丽娟.聚乙烯醇生物降解性能的研究[D].南京:南京理工大学, 2005.
[16]Lopez BLO, Mejia AIG, Sierra LG. Biodegradability of poly(vinyl alcohol). Polymer Engineering&Science[J]. 2010, 39(8):1346-1352.
[17]Choi K, Park C, Kim S, et al. Polyvinyl alcohol degradation by microbacterium barkeri KCCM 10507 and Paenibacillus amylolyticus KCCM 10508 in dyeing wastewater[J]. Journal of Microbiology&Biotechnology, 2004, 14(5):1009-1013.
[18]Mori T, Sakimoto M, Kagi TT. Isolation and characterization of a strain of bacillus megaterium that degrades poly(vinyl alcohol)[J]. Bioscience Biotechnology&Biochemistry, 1996,60(2):330-332.
[19]Chen J, Zhang Y, Du GC, et al. Biodegradation of polyvinyl alcohol by a mixed microbial culture[J]. Enzymeµbial technology, 2007, 40(7):1686-1691.
[20]赵一凡.聚乙烯醇降解菌的筛选及其关键酶的应用研究[D].无锡:江南大学, 2017.
[21]张颖.微生物降解聚乙烯醇的条件及降解过程机制研究[D].无锡:江南大学, 2006.
[22]厉成宣,范雪荣,王强,等.退浆废水中PVA对环境的影响及其降解性能[J].印染助剂, 2007, 24(6):7-10.