Preparation and Characterization of Chitinase from Marine Bacteria <i>Aeromonas</i> sp. YS-54

CHEN Sunan; LENG Kailiang; YAN Mingyan; LI Yinping; YU Yuan

doi:10.13386/j.issn1002-0306.2023090122

Volume 45 Issue 16

Aug. 2024

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Science and Technology of Food Industry > 2024 > 45(16): 182-190. > DOI: 10.13386/j.issn1002-0306.2023090122

CHEN Sunan, LENG Kailiang, YAN Mingyan, et al. Preparation and Characterization of Chitinase from Marine Bacteria Aeromonas sp. YS-54[J]. Science and Technology of Food Industry, 2024, 45(16): 182−190. (in Chinese with English abstract). doi: 10.13386/j.issn1002-0306.2023090122.

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Preparation and Characterization of Chitinase from Marine Bacteria Aeromonas sp. YS-54

1.
College of Marine Science and Biological Engineering, Qingdao University of Science and Technology,Qingdao 266042, China
2.
Key Laboratory of Sustainable Development of Polar Fishery, Ministry of Agriculture and Rural Affairs, Yellow Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, Qingdao 266071, China

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Received Date: September 12, 2023
Available Online: June 19, 2024

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Abstract

Abstract

To achieve efficient degradation of crustacean raw materials and eco-preparation of N-acetyl-oligosaccharides, YS-54 strain screened from offshore soil in Qingdao was used for chitinase preparation in the present study. Chitinase was prepared through 60% ammonium sulfate precipitation and Sephadex G-100 gel filtration chromatography. Enzymatic properties such as optimal temperature, optimal pH, and substrate specificity were characterized. Mass spectrometry was used to determine the polymerization degree of enzymatic products. The 16S rRNA identification result showed that YS-54 strain belonged to the Aeromonas species. Chitinase from Aeromonas sp. YS-54 showed a specific activity of 23.44 U/mg. The molecular weights of chitinase were 63 and 75 kDa as shown by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). Chitinase was stable under the condition of 40 ℃ and pH5.0. The enzymatic activity was significantly activated by Ba²⁺, Co²⁺, Mg²⁺ and TritonX-100 (P<0.05), while inhibited by Fe³⁺, Cu²⁺ and SDS. The specific activity of chitinase toward α-chitin was 7.99 U/mg, 34.09% of that toward colloidal chitin. The polymerization degree of enzymatic hydrolysis products from colloidal chitin and α-chitin were 1~4 and 1~3, respectively. Chitinase from Aeromonas sp. YS-54 was stable in broad range of temperature and pH, meanwhile showed high catalytic efficiency toward α-chitin. The hydrolysis characteristics of chitinase from Aeromonas sp. YS-54 would provide technical support for the comprehensive utilization of crustacean waste.
- chitinase,
- enzymatic property,
- substrate specificity,
- N-acetyl-oligosaccharides,
- marine bacteria Aeromonas sp.

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References

[1]	刘宇, 方国宏, 戎素红, 等. 虾、蟹壳利用的研究进展[J]. 食品安全质量检测学报,2018,9(3):461−466. [LIU Y, FANG G H, RONG S H, et al. Research progress on the utilization of shrimp and crab shells[J]. Journal of Food Safety and Quality,2018,9(3):461−466.] doi: 10.3969/j.issn.2095-0381.2018.03.001 LIU Y, FANG G H, RONG S H, et al. Research progress on the utilization of shrimp and crab shells[J]. Journal of Food Safety and Quality, 2018, 9（3）: 461−466. doi: 10.3969/j.issn.2095-0381.2018.03.001
[2]	ZHANG A L, HE Y M, WEI G G, et al. Molecular characterization of a novel chitinase CmChi1 from Chitinolyticbacter meiyuanensis SYBC-H1 and its use in N-acetyl-D-glucosamine production[J]. Biotechnology for Biofuels,2018,11:179. doi: 10.1186/s13068-018-1169-x
[3]	RAHAYU A P, ISLAMI A F, SAPUTRA E, et al. The impact of the different types of acid solution on the extraction and adsorption performance of chitin from shrimp shell waste[J]. International Journal of Biological Macromolecules,2022,194:843−850. doi: 10.1016/j.ijbiomac.2021.11.137
[4]	SEBASTIAN J, ROUISSI T, BRAR S K, et al. Microwave-assisted extraction of chitosan from Rhizopus oryzae NRRL 1526 biomass[J]. Carbohydrate Polymers,2019,219:431−440. doi: 10.1016/j.carbpol.2019.05.047
[5]	LUCAS A J D S, ORESTE E Q, COSTA H L G, et al. Extraction, physicochemical characterization, and morphological properties of chitin and chitosan from cuticles of edible insects[J]. Food Chemistry,2021,343:128550. doi: 10.1016/j.foodchem.2020.128550
[6]	RANI T S, MADHUPRAKASH J, PODILE A R. Chitinase-E from Chitiniphilus shinanonensis generates chitobiose from chitin flakes[J]. International Journal of Biological Macromolecules,2020,163:1037−1043. doi: 10.1016/j.ijbiomac.2020.07.052
[7]	刘力睿, 潘杰, 李猛, 等. 微生物几丁质酶的研究进展、应用及展望[J]. 生物资源,2020,42(5):494−504. [LIU L R, PAN J, LI M, et al. Research progress, application and prospects of microbial chitinase[J]. Biotic Resources,2020,42(5):494−504.] LIU L R, PAN J, LI M, et al. Research progress, application and prospects of microbial chitinase[J]. Biotic Resources, 2020, 42（5）: 494−504.
[8]	PATEL S, GOYAL A. Chitin and chitinase:Role in pathogenicity, allergenicity and health[J]. International Journal of Biological Macromolecules,2017,97:331−338. doi: 10.1016/j.ijbiomac.2017.01.042
[9]	MA X, GÖZAYDIN G, YANG H, et al. Upcycling chitin-containing waste into organonitrogen chemicals via an integrated process[J]. Proceedings of the National Academy of Sciences of the United States of America,2020,117(14):7719−7728.
[10]	赵沙, 颜子娟, 张舒, 等. 细菌几丁质酶结构、功能及分子设计的研究进展[J]. 生物化学与生物物理进展,2022,49(7):1179−1191. [ZHAO S, YAN Z J, ZHANG S, et al. Research progress on structure, function and molecular design of bacterial chitinase[J]. Progress in Biochemistry and Biophysics,2022,49(7):1179−1191.] ZHAO S, YAN Z J, ZHANG S, et al. Research progress on structure, function and molecular design of bacterial chitinase[J]. Progress in Biochemistry and Biophysics, 2022, 49（7）: 1179−1191.
[11]	周玉玲, 蒋思婧, 贺妮莎, 等. 微生物几丁质酶研究进展及其在N-乙酰氨基葡萄糖制备中的应用[J]. 微生物学报,2021,61(8):2192−2204. [ZHOU Y L, JIANG S J, HE N S, et al. Research progress of microbial chitinase and its application in the preparation of N-acetylglucosamine[J]. Acta Microbiologica Sinica,2021,61(8):2192−2204.] ZHOU Y L, JIANG S J, HE N S, et al. Research progress of microbial chitinase and its application in the preparation of N-acetylglucosamine[J]. Acta Microbiologica Sinica, 2021, 61（8）: 2192−2204.
[12]	LÜ J R, ZHANG Y M, MA M H, et al. Characterization of chitinase from Exiguobacterium antarcticum and its bioconversion of crayfish shell into chitin oligosaccharides[J]. Food Research International,2022,158:111517. doi: 10.1016/j.foodres.2022.111517
[13]	高兆建, 丁飞鸿, 陈欢, 等. 产黄青霉抗真菌几丁质酶的纯化及特性分析[J]. 食品科学,2021,42(14):129−136. [GAO Z J, DING F H, CHEN H, et al. Purification and characterization of antifungal chitinase from Penicillium chrysogenum[J]. Food Science,2021,42(14):129−136.] doi: 10.7506/spkx1002-6630-20200229-329 GAO Z J, DING F H, CHEN H, et al. Purification and characterization of antifungal chitinase from Penicillium chrysogenum[J]. Food Science, 2021, 42（14）: 129−136. doi: 10.7506/spkx1002-6630-20200229-329
[14]	CHEN W, JIANG X, YANG Q. Glycoside hydrolase family 18 chitinases:The known and the unknown[J]. Biotechnology Advances,2020,43:107553. doi: 10.1016/j.biotechadv.2020.107553
[15]	OLIVERIRA S T, AZEVEDO M I G, CUNHA R M S, et al. Structural and functional features of a class VI chitinase from cashew (Anacardium occidentale L.) with antifungal properties[J]. Phytochemistry,2020,180:112527. doi: 10.1016/j.phytochem.2020.112527
[16]	HALDER S K, MAITY C, JANA A, et al. Chitinases biosynthesis by immobilized Aeromonas hydrophila SBK1 by prawn shells valorization and application of enzyme cocktail for fungal protoplast preparation[J]. Journal of Bioscience and Bioengineering,2014,117(2):170−177. doi: 10.1016/j.jbiosc.2013.07.011
[17]	AKEED Y, ATRASH F, NAFFAA W. Partial purification and characterization of chitinase produced by Bacillus licheniformis B307[J]. Heliyon,2020,6(5):E03858. doi: 10.1016/j.heliyon.2020.e03858
[18]	BHUVANACHANDRA B, PODILE A R. A transglycosylating chitinase from Chitiniphilus shinanonensis (CsChiL) hydrolyzes chitin in a processive manner[J]. International Journal of Biological Macromolecules,2020,145:1−10. doi: 10.1016/j.ijbiomac.2019.12.134
[19]	FU X, GUO Y X, JIN Y G, et al. Bioconversion of chitin waste using a cold-adapted chitinase to produce chitin oligosaccharides[J]. LWT,2020,133:109863. doi: 10.1016/j.lwt.2020.109863
[20]	YANO S, KANNO H, TSUHAKO H, et al. Cloning, expression, and characterization of a GH 19-type chitinase with antifungal activity from Lysobacter sp. MK9-1[J]. Journal of Bioscience and Bioengineering,2021,131(4):348−355. doi: 10.1016/j.jbiosc.2020.11.005
[21]	LEE Y S, KIM K Y. Statistical optimization of medium components for chitinase production by Pseudomonas fluorescens strain HN1205:role of chitinase on egg hatching inhibition of root-knot nematode[J]. Biotechnology & Biotechnological Equipment,2015,29(3):470−478.
[22]	WANG S L, LIN C L, LIANG T W, et al. Conversion of squid pen by Serratia ureilytica for the production of enzymes and antioxidants[J]. Bioresource Technology,2009,100(1):316−323. doi: 10.1016/j.biortech.2008.06.026
[23]	HALDER S K, JANA A, PAUL T et al. Purification and biochemical characterization of chitinase of Aeromonas hydrophila SBK1 biosynthesized using crustacean shell[J]. Biocatalysis and Agricultural Biotechnology,2016,5:211−218. doi: 10.1016/j.bcab.2015.11.003
[24]	HALDER S K, MAITY C, JANA A, et al. Proficient biodegradation of shrimp shell waste by Aeromonas hydrophila SBK1 for the concomitant production of antifungal chitinase and antioxidant chitosaccharides[J]. International Biodeterioration & Biodegradation,2013,79:88−97.
[25]	WANG C Y, CHEN X M, ZHOU N, et al. Property and function of a novel chitinase containing dual catalytic domains capable of converting chitin into N-acetyl-D-glucosamine[J]. Frontiers in Microbiology,2022,13:790301. doi: 10.3389/fmicb.2022.790301
[26]	WU Y L, WANG S, YANG D F, et al. The discovery, enzymatic characterization and functional analysis of a newly isolated chitinase from marine-derived fungus Aspergillus fumigatus df347[J]. Marine Drugs,2022,20(8):520. doi: 10.3390/md20080520
[27]	BHVANACHANDRA B, SIVARAMAKRISHNA D, ALIM S, et al. Deciphering the thermotolerance of chitinase O from Chitiniphilus shinanonensis by in vitro and in silico studies[J]. International Journal of Biological Macromolecules,2022,210:44−52. doi: 10.1016/j.ijbiomac.2022.05.013
[28]	ZHANG W J, MA J W, YAN Q J, et al. Biochemical characterization of a novel acidic chitinase with antifungal activity from Paenibacillus xylanexedens Z2-4[J]. International Journal of Biological Macromolecules,2021,182:1528−1536. doi: 10.1016/j.ijbiomac.2021.05.111
[29]	SU H P, GAO L, SUN J N, et al. Engineering a carbohydrate binding module to enhance chitinase catalytic efficiency on insoluble chitinous substrate[J]. Food Chemistry,2021,355:129462. doi: 10.1016/j.foodchem.2021.129462
[30]	王燕, 赵力超, 陈洁兰, 等. 菠萝蛋白酶工业化提取工艺的改良及酶学性质研究[J]. 食品工业科技,2011,32(10):353−357. [WANG Y, ZHAO L C, CHEN J L, et al. Study on improvement of industrial extraction of bromelain and its enzymology characterization[J]. Science and Technology of Food Industry,2011,32(10):353−357.] WANG Y, ZHAO L C, CHEN J L, et al. Study on improvement of industrial extraction of bromelain and its enzymology characterization[J]. Science and Technology of Food Industry, 2011, 32（10）: 353−357.
[31]	陈茜文, 王佳丽, 许春雨. 克氏原螯虾壳膜几丁质酶的分离纯化[J]. 食品工业科技,2018,39(23):159−163. [CHEN Q W, WANG J L, XU C Y. Isolation and purification of chitinase from shellfish of Procambarus clarkia[J]. Science and Technology of Food Industry,2018,39(23):159−163.] CHEN Q W, WANG J L, XU C Y. Isolation and purification of chitinase from shellfish of Procambarus clarkia[J]. Science and Technology of Food Industry, 2018, 39（23）: 159−163.
[32]	徐勤茜, 朱国威, 赵梓伶, 等. 酸橙内生菌Bacillus thuringiensis Bt028几丁质酶的分离纯化及其酶学性质[J]. 食品工业科技,2022,43(11):159−166. [XU Q Q, ZHU G W, ZHAO Z L, et al. Separation, purification and characterization of chitinase of the endophytic bacterium Bacillus thuringiensis Bt028 isolated from sour orange[J]. Science and Technology of Food Industry,2022,43(11):159−166.] XU Q Q, ZHU G W, ZHAO Z L, et al. Separation, purification and characterization of chitinase of the endophytic bacterium Bacillus thuringiensis Bt028 isolated from sour orange[J]. Science and Technology of Food Industry, 2022, 43（11）: 159−166.
[33]	QIU S T, ZHOU S P, TAN Y, et al. Biodegradation and prospect of polysaccharide from crustaceans[J]. Marine Drugs,2022,20(5):310. doi: 10.3390/md20050310
[34]	宋阳, 陈梦, 吴新财, 等. 产几丁质酶菌株Acinetobacter sp. CZW011的筛选鉴定与酶学性质研究[J]. 中国食品添加剂,2022,33(11):1−8. [SONG Y, CHEN M, WU X C, et al. Screening, identification and enzymatic properties of Acinetobacter sp. CZW011 strain produced by chitinase[J]. China Food Additives,2022,33(11):1−8.] SONG Y, CHEN M, WU X C, et al. Screening, identification and enzymatic properties of Acinetobacter sp. CZW011 strain produced by chitinase[J]. China Food Additives, 2022, 33（11）: 1−8.
[35]	严佳佳, 尤田, 张雪梅, 等. 一种海洋几丁质酶产生菌的筛选及酶学性质研究[J]. 食品与发酵工业,2023,49(13):40−48. [YAN J J, YOU T, ZHANG X M, et al. Screening of a marine chitinase-producing bacterium and its enzymatic properties[J]. Food and Fermentation Industry,2023,49(13):40−48.] YAN J J, YOU T, ZHANG X M, et al. Screening of a marine chitinase-producing bacterium and its enzymatic properties[J]. Food and Fermentation Industry, 2023, 49（13）: 40−48.
[36]	MATHEW G, MADHAVAN A, ARUN K B, et al. Thermophilic chitinases:Structural, functional and engineering attributes for industrial applications[J]. Applied Biochemistry and Biotechnology,2021,193:142−164. doi: 10.1007/s12010-020-03416-5
[37]	HAO Z K, WU H G, YANG M L, et al. Cloning, expression and 3D structure prediction of chitinase from Chitinolyticbacter meiyuanensis SYBC-H1[J]. International Journal of Molecular Sciences,2016,17:825. doi: 10.3390/ijms17060825
[38]	YU G Q, LIU G H, LIU T Y, et al. Activities of family 18 chitinases on amorphous regenerated chitin thin films and dissolved chitin oligosaccharides:Comparison with family 19 chitinases[J]. Biomacromolecules 2023, 24(2):566−575.
[39]	BHUSHAN B, HOONDAL G S. Isolation, purification and properties of a thermostable chitinase from an alkalophilic Bacillus sp. BG-11[J]. Biotechnology Letters,1998,20:157−159. doi: 10.1023/A:1005328508227
[40]	ZHANG A L, MO X F, ZHOU N, et al. A novel bacterial β-N-acetyl glucosaminidase from Chitinolyticbacter meiyuanensis possessing transglycosylation and reverse hydrolysis activities[J]. Biotechnology for Biofuels,2020,13:115. doi: 10.1186/s13068-020-01754-4
[41]	SUN B, ZHAO X C, XU B R, et al. Discovering and designing a chimeric hyperthermophilic chitinase for crystalline chitin degradation[J]. ACS Sustainable Chemistry & Engineering,2023,11(12):4690−4698.
[42]	KITAOKU Y, TAIRA T, NUMATA T, et al. Structure, mechanism, and phylogeny of LysM-chitinase conjugates specifically found in fern plants[J]. Plant Science,2022,321:111310. doi: 10.1016/j.plantsci.2022.111310
[43]	MATROODI S, MOTALLEBI M, ZAMANI M, et al. Designing a new chitinase with more chitin binding and antifungal activity[J]. World Journal of Microbiology & Biotechnology,2013,29:1517−1523.
[44]	LÜ C Y, GU T Y, MA R, et al. Biochemical characterization of a GH19 chitinase from Streptomyces alfalfae and its applications in crystalline chitin conversion and biocontrol[J]. International Journal of Biological Macromolecules,2021,167:193−201. doi: 10.1016/j.ijbiomac.2020.11.178

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