Production of Pectinolytic Enzymes by Two Bacillus spp. Strains and Their Application in Flax Degumming
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摘要
This study demonstrated that Bacillus licheniformis HDYM-03 and Bacillus megaterium HDYM-09, isolated from a liquid sample of flax retting pool, were able to produce pectinolytic enzymes using polysaccharides as substrates. Bacillus megaterium HDYM-09 produced pectin lyase that exhibited the highest activity of 2116.71 ± 11.55 U/mL. Bacillus licheniformis HDYM-03 produced pectate lyase that exhibited the highest activity of 611.21 ± 14.54 U/mL. Based on these findings, we constructed four retting systems to degrade the pectin substance. The results showed that the content of galacturonic acid in the mixed system was 529.21 μg/mL, the content of reducing sugar was 98.14 mg/mL, and the weight loss ratio of cells reached 19.49%, which were significantly higher than those in other systems. The mixed system has more advantages, and the utilization rate of degumming was higher, which further ensured that the degumming can be carried out efficiently and quickly. The mixed system exhibits feasible applications in the fiber and textile industry.
This study demonstrated that Bacillus licheniformis HDYM-03 and Bacillus megaterium HDYM-09, isolated from a liquid sample of flax retting pool, were able to produce pectinolytic enzymes using polysaccharides as substrates. Bacillus megaterium HDYM-09 produced pectin lyase that exhibited the highest activity of 2116.71 ± 11.55 U/mL. Bacillus licheniformis HDYM-03 produced pectate lyase that exhibited the highest activity of 611.21 ± 14.54 U/mL. Based on these findings, we constructed four retting systems to degrade the pectin substance. The results showed that the content of galacturonic acid in the mixed system was 529.21 μg/mL, the content of reducing sugar was 98.14 mg/mL, and the weight loss ratio of cells reached 19.49%, which were significantly higher than those in other systems. The mixed system has more advantages, and the utilization rate of degumming was higher, which further ensured that the degumming can be carried out efficiently and quickly. The mixed system exhibits feasible applications in the fiber and textile industry.
This study demonstrated that Bacillus licheniformis HDYM-03 and Bacillus megaterium HDYM-09, isolated from a liquid sample of flax retting pool, were able to produce pectinolytic enzymes using polysaccharides as substrates. Bacillus megaterium HDYM-09 produced pectin lyase that exhibited the highest activity of 2116.71 ± 11.55 U/mL. Bacillus licheniformis HDYM-03 produced pectate lyase that exhibited the highest activity of 611.21 ± 14.54 U/mL. Based on these findings, we constructed four retting systems to degrade the pectin substance. The results showed that the content of galacturonic acid in the mixed system was 529.21 μg/mL, the content of reducing sugar was 98.14 mg/mL, and the weight loss ratio of cells reached 19.49%, which were significantly higher than those in other systems. The mixed system has more advantages, and the utilization rate of degumming was higher, which further ensured that the degumming can be carried out efficiently and quickly. The mixed system exhibits feasible applications in the fiber and textile industry.
引文
1.Foulk JA,Rho D,Alcock MM et al(2011)Modifi cations caused by enzyme-retting and their effect on composite performance.Adv Mater Sci Eng 2011:179023
2.Akin DE(2013)Linen most useful:perspectives on str ucture,chemistry,and enzymes for retting flax.ISRN Biotech2013:186534
3.Bai YK,Liu BY,He LF(2006)Pectinase and its applications in pulp and paper industry.Trans China Pulp Paper 21(2):88-91(in Chinese)
4.Deng WK,Liu EP,Guo AP et al(2009)Study on characteristics of pectinase from a bacteria for degumming pineapple leaf fi ber.Guangdong Agric Sci 9(6):190-193(in Chinese)
5.Zheng K,Liu ZC,Duan SW et al(2012)Mechanism of the pectinases used for bast fi bre bio-degumming.Curr Biotech2(6):404-410(in Chinese)
6.Huang H,Zhang X(2005)Effects of buffer system on laccase determination.Ind Microbiol 35(4):29-32(in Chinese)
7.Wu CB,Cao YB,Wu HQ et al(2010)The application for Bacillus subtilis HW201 in flax retting-intermediate test.Plant Fiber Sci China 32(6):327-330(in Chinese)
8.Ge JP,Ling HZ,Song G et al(2004)The study on isolation of pectinase-producing strains and its cultural conditions.China Biotechnol 24(8):93-95(in Chinese)
9.Li ZM,Zhang HX,Bai ZH et al(2010)Advance in microbial pectinases research.Biotechnol Bull 3(1):42-49(in Chinese)
10.Wang HL,Li XM,Ma YH et al(2014)Characterization and high-level expression of a metagenome-derived alkaline pectate lyase in recombinant Escherichia coli.Process Biochem49(1):69-76
11.Zhuge B,Du GC,Zhuge J et al(2008)Properties of alkaline pectate lyase from recombinant strain E.coli JM109.Acta Microbiol Sin 48(1):121-125(in Chinese)
12.Zhao D,Liu P,Pan C et al(2016)Flax retting by degumming composite enzyme produced by Bacillus licheniformis HDYM-04 and effect on fi ber properties.J Text Inst 108(4):507-510
13.Ding J,Zhang X,Yao X et al(2010)Determination of sisal pectin content by carbazole colorimetric method.Food Res Dev31(11):138-140(in Chinese)
14.Wang J,Nie G,Li S et al(2010)Optimal wavelength for determining the content of reducing sugar by DNS method.Henan Agric Sci 4(1):115-118(in Chinese)
15.Wang T,Qiu H,Chen H et al(2008)Exploration of the determination conditions of the content of D-galacturonic acid with the method of 3,5-dinitrosalicylic acid(DNS).J Anhui Agric Sci 36(16):6829-6832(in Chinese)
16.Soriano M,Diaz P,Pastor F(2005)Pectinolytic systems of two aerobic sporogenous bacterial strains with high activity on pectin.Curr Microbiol 50(2):114-118
17.Doyle EA,Lambert KN(2002)Cloning and characterization of an esophageal-gland-specifi c pectate lyase from the root-knot nematode Meloidogyne javanica.Mol Plant Microbe Interact15(6):549-554
18.Wang Q,Chen A,Fang G et al(2017)Isolation of Bacillus cereus P05 and Pseudomonas sp.X12 and their application in the ramie retting.Ind Crops Prod 97(3):518-524
19.Sharma N,Rathore M,Sharma M(2012)Microbial pectinase:sources,characterization and applications.Rev Environ Sci Bio12(1):45-60
20.Chiliveri SR,Koti S,Linga VR(2016)Retting and degumming of natural fi bers by pectinolytic enzymes produced from Bacillus tequilensis SV11-UV37 using solid state fermentation.SpringerPlus 5:559
21.Biswapriya D,Kalyan C,Sagarmoy G et al(2012)Effect of efficient pectinolytic bacterial isolates on retting and fi bre quality of jute.Ind Crops Prod 36(1):415-419
22.Zhao D,Liu PF,Pan C et al(2016)Bacterial succession and metabolite changes during flax(Linum usitatissimum L.)retting with Bacillus cereus HDYM-02.Sci Rep 6:31812
23.Di Candilo M,Bonatti PM,Guidetti C et al(2010)Effects of selected pectinolytic bacterial strains on water-retting of hemp and fi bre properties.J Appl Microbiol 108(1):194-203
24.Mukhopadhyay A,Dutta N,Dhrubajyoti et al(2013)Degumming of ramie fi ber and the production of reducing sugars from waste peels using nanoparticle supplemented pectate lyase.Bioresour Technol 137:202-208
25.Yu W,Wang G,Han J et al(2012)Application of functional degumming bacteria in hemp retting.Hubei Agric Sci 51(21):4772-4775(in Chinese)
26.Brühlmann F,Kim KS,Zimmerman W et al(1994)Pectinolytic enzymes from Actinomycetes for the degumming of ramie bast fi bers.Appl Environ Microbiol 60(6):7-12
2.Akin DE(2013)Linen most useful:perspectives on str ucture,chemistry,and enzymes for retting flax.ISRN Biotech2013:186534
3.Bai YK,Liu BY,He LF(2006)Pectinase and its applications in pulp and paper industry.Trans China Pulp Paper 21(2):88-91(in Chinese)
4.Deng WK,Liu EP,Guo AP et al(2009)Study on characteristics of pectinase from a bacteria for degumming pineapple leaf fi ber.Guangdong Agric Sci 9(6):190-193(in Chinese)
5.Zheng K,Liu ZC,Duan SW et al(2012)Mechanism of the pectinases used for bast fi bre bio-degumming.Curr Biotech2(6):404-410(in Chinese)
6.Huang H,Zhang X(2005)Effects of buffer system on laccase determination.Ind Microbiol 35(4):29-32(in Chinese)
7.Wu CB,Cao YB,Wu HQ et al(2010)The application for Bacillus subtilis HW201 in flax retting-intermediate test.Plant Fiber Sci China 32(6):327-330(in Chinese)
8.Ge JP,Ling HZ,Song G et al(2004)The study on isolation of pectinase-producing strains and its cultural conditions.China Biotechnol 24(8):93-95(in Chinese)
9.Li ZM,Zhang HX,Bai ZH et al(2010)Advance in microbial pectinases research.Biotechnol Bull 3(1):42-49(in Chinese)
10.Wang HL,Li XM,Ma YH et al(2014)Characterization and high-level expression of a metagenome-derived alkaline pectate lyase in recombinant Escherichia coli.Process Biochem49(1):69-76
11.Zhuge B,Du GC,Zhuge J et al(2008)Properties of alkaline pectate lyase from recombinant strain E.coli JM109.Acta Microbiol Sin 48(1):121-125(in Chinese)
12.Zhao D,Liu P,Pan C et al(2016)Flax retting by degumming composite enzyme produced by Bacillus licheniformis HDYM-04 and effect on fi ber properties.J Text Inst 108(4):507-510
13.Ding J,Zhang X,Yao X et al(2010)Determination of sisal pectin content by carbazole colorimetric method.Food Res Dev31(11):138-140(in Chinese)
14.Wang J,Nie G,Li S et al(2010)Optimal wavelength for determining the content of reducing sugar by DNS method.Henan Agric Sci 4(1):115-118(in Chinese)
15.Wang T,Qiu H,Chen H et al(2008)Exploration of the determination conditions of the content of D-galacturonic acid with the method of 3,5-dinitrosalicylic acid(DNS).J Anhui Agric Sci 36(16):6829-6832(in Chinese)
16.Soriano M,Diaz P,Pastor F(2005)Pectinolytic systems of two aerobic sporogenous bacterial strains with high activity on pectin.Curr Microbiol 50(2):114-118
17.Doyle EA,Lambert KN(2002)Cloning and characterization of an esophageal-gland-specifi c pectate lyase from the root-knot nematode Meloidogyne javanica.Mol Plant Microbe Interact15(6):549-554
18.Wang Q,Chen A,Fang G et al(2017)Isolation of Bacillus cereus P05 and Pseudomonas sp.X12 and their application in the ramie retting.Ind Crops Prod 97(3):518-524
19.Sharma N,Rathore M,Sharma M(2012)Microbial pectinase:sources,characterization and applications.Rev Environ Sci Bio12(1):45-60
20.Chiliveri SR,Koti S,Linga VR(2016)Retting and degumming of natural fi bers by pectinolytic enzymes produced from Bacillus tequilensis SV11-UV37 using solid state fermentation.SpringerPlus 5:559
21.Biswapriya D,Kalyan C,Sagarmoy G et al(2012)Effect of efficient pectinolytic bacterial isolates on retting and fi bre quality of jute.Ind Crops Prod 36(1):415-419
22.Zhao D,Liu PF,Pan C et al(2016)Bacterial succession and metabolite changes during flax(Linum usitatissimum L.)retting with Bacillus cereus HDYM-02.Sci Rep 6:31812
23.Di Candilo M,Bonatti PM,Guidetti C et al(2010)Effects of selected pectinolytic bacterial strains on water-retting of hemp and fi bre properties.J Appl Microbiol 108(1):194-203
24.Mukhopadhyay A,Dutta N,Dhrubajyoti et al(2013)Degumming of ramie fi ber and the production of reducing sugars from waste peels using nanoparticle supplemented pectate lyase.Bioresour Technol 137:202-208
25.Yu W,Wang G,Han J et al(2012)Application of functional degumming bacteria in hemp retting.Hubei Agric Sci 51(21):4772-4775(in Chinese)
26.Brühlmann F,Kim KS,Zimmerman W et al(1994)Pectinolytic enzymes from Actinomycetes for the degumming of ramie bast fi bers.Appl Environ Microbiol 60(6):7-12