Immobilization of Marine Urease and Its Utilization in the Treatment of Urea Wastewater

doi:10.13560/j.cnki.biotech.bull.1985.2019-0254

Abstract

Abstract: The objective is to develop an appropriate immobilization strategy for marine alkaline urease and to provide technical assessments for urea wastewater treatment via the immobilized urease. The enzyme was immobilized by 3 kinds of materials,and the immobilization effects,enzyme properties were examined. The immobilized urease was loaded in a column for urea wastewater treatment and the operating parameters were optimized. Urease immobilized by alginate-CaCl₂ was the best with an immobilization rate reaching 61.47%. Comparing to the free urease with optimal temperature 55℃ and pH 8.5,the immobilized urease increased to 70℃,and pH 9.5,respectively,and sustained over 90% activity after incubating at 35-85℃ for 30 min or at pH 7-10.5 for 6 h. Furthermore,its half-life exceeded 3 months when stored at 4℃. The treatment apparatus achieved a flow rate of 19 mL/min at 70℃ with an initial urea decomposition rate of 90.25%,and the efficiency remained constant for 35 days. In conclusion,the optimal temperature and pH,the thermostability and the alkali resistance of the marine urease are much improved after being immobilized by alginate-CaCl₂. The urea wastewater treatment applying the immobilized marine urease represents a considerable alternative technique besides chemical method.

Key words: marine urease, immobilization, urea, wastewater treatment

LIU Chang-rong, ZHANG Feng-li, LI Zhi-yong. Immobilization of Marine Urease and Its Utilization in the Treatment of Urea Wastewater[J]. Biotechnology Bulletin, 2019, 35(9): 75-82.

References

[1] Kafarski P, Talma M.Recent advances in design of new urease inhibitors:A review[J]. J Adv Res, 2018, 13:101-112.
[2] Krajewska B, Ureases I.Functional, catalytic and kinetic properties: A review[J]. J Mol Catalysis B Enzym, 2009, 59(1-3):9-21.
[3] Qin Y, Cabral JMS.Review properties and applications of urease[J]. Biocatalysis and Biotransformation, 2002, 20(1):1-14.
[4] Heffer P, Prud'homme M. Short-term fertilizer outlook 2018-2019[C]. Beijing:International Fertilizer Association Strategic Forum, 2018.
[5] Latkar M, Chakrabarti T.Performance of upflow anaerobic sludge blanket reactor carrying out biological hydrolysis of urea[J]. Water Environment Research, 1994. 66(1):12-15.
[6] 乔德阳. 利用固定化脲酶处理尿素废水的研究[J]. 徐州师范大学学报:自然科学版, 2001, 19(2):47-49.
[7] Zeinali M, Lenjannezhadian H.Degradation of urea by entrapped cross-linked urease aggregates:a combinatorial approach to urease stabilization for environmental and industrial applications[J]. Int J Environ Sci Technol, 2018, 15(1):49-56.
[8] Krajewska B. Ureases. II.Properties and their customizing by enzyme immobilizations:A review[J]. J Mol Catalysis B Enzyme, 2009, 59(1-3):22-40.
[9] 张莉娟. 脲酶的组合固定化研究[D]. 武汉:武汉理工大学, 2012.
[10] Jahns T.Purification and properties of urease from Sporobolomyces roseus[J]. Antonie Van Leeuwenhoek, 1995, 68(3):209-214.
[11] Palinska KA, Jahns T, Rippka R, et al.Prochlorococcus marinus strain PCC 9511, a picoplanktonic cyanobacterium, synthesizes the smallest urease[J]. Microbiology, 2000, 146(12):3099-3107.
[12] Phillips A, Pretorius GH, Du Toit PJ.A survey of yeast ureases and characterisation of partially purified Rhodosporidium paludigenum urease[J]. FEMS Microbiology Letters, 1991, 79(1):21-25.
[13] Tanriseven A, Dogan S.Immobilization of invertase within calcium alginate gel capsules[J]. Process Biochemistry, 2001, 36(11):1081-1083.
[14] Srivastava PK, Kayastha AM, Srinivasan. Characterization of gelatin-immobilized pigeonpea urease and preparation of a new urea biosensor[J]. Biotechnol Appl Biochem, 2001, 34(1):55-62.
[15] Liu Q, Chen Y, Yuan M, et al.A Bacillus paralicheniformis iron-containing urease reduces urea concentrations in rice wine[J]. Appl Environ Microbiol, 2017, 83(17):e01258-17.
[16] 段晓琛, 盛军, 徐甲坤, 等. 海洋脂肪酶ADM47601固定化方法的研究[J]. 海洋与湖沼, 2013, 44(5):1311-1317.
[17] Moreira SM, Moreira-Santos M, Lucia G, et al.Immobilization of the marine microalga Phaeodactylum tricornutum in alginate for in situ experiments:Bead stability and suitability[J]. Enzyme and Microbial Technology, 2006, 38(1-2):135-141.
[18] 莫旖, 姬琳, 柳畅先. 固定化脲酶的制备及应用[J]. 分析科学学报, 2013, 29(1):85-88.
[19] 骆东辉, 刘安妮, 李明芳, 等. ¹³¹I标记海藻酸钠包裹明胶微球的制备及性能评价[J]. 高等学校化学学报, 2011, 32(6):1412-1417.
[20] Tiwari A, Aryal S, Pilla S, et al.An amperometric urea biosensor based on covalently immobilized urease on an electrode made of hyperbranched polyester functionalized gold nanoparticles[J]. Talanta, 2009, 78(4-5):1401-1407.
[21] Yang L, Liu X, Zhou N, et al.Characteristics of refold acid urease immobilized covalently by graphene oxide-chitosan composite beads[J]. J Biosci Bioeng, 2019, 127(1):16-22.
[22] Kayastha AM, Srivastava PK.Pigeonpea(Cajanus cajan L.)urease immobilized on glutaraldehyde-activated chitosan beads and its analytical applications[J]. Applied Biochemistry and Biotechnology, 2001, 96(1-3):41-53.
[23] Kirdeciler SK, Soy E, Ozturk S, et al.A novel urea conductometric biosensor based on zeolite immobilized urease[J]. Talanta, 2011, 85(3):1435-1441.
[24] Das N, Kayastha AM, Malhotra OP.Immobilization of urease from pigeonpea(Cajanus cajan L.)in polyacrylamide gels and calcium alginate beads[J]. Biotechnol Appl Biochem, 1998, 1:25-29.
[25] Prakash O, Upadhyay LSB.Physico-chemical characterization of watermelon urease upon immobilization in alginate beads[J]. J Plant Biochem Biotechnol, 2005, 14(2):209-213.
[26] Kutcherlapati SN, Yeole N, Jana T.Urease immobilized polymer hydrogel:Long-term stability and enhancement of enzymatic activity[J]. J Colloid Interface Sci, 2016, 463:164-172.
[27] Alatawi FS, Monier M, Elsayed NH.Amino functionalization of carboxymethyl cellulose for efficient immobilization of urease[J]. Int J Biol Macromol, 2018, 114:1018-1025.
[28] Zhang L, Du Y, Song JY, et al.Biocompatible magnetic nanoparticles grafted by poly(carboxybetaine acrylamide)for enzyme immobilization[J]. International Journal of Biological Macromolecules, 2018, 118:1004-1012.
[29] Monier M, EI-Sokkary AMA.Modification and characterization of cellulosic cotton fibers for efficient immobilization of urease[J]. Int J Biol Macromol, 2012, 51(1-2):18-24.
[30] Dogac YI, Deveci I, Teke M, et al.TiO₂ beads and TiO₂-chitosan beads for urease immobilization[J]. Materials Science and Engineering:C, 2014, 42:429-435.