诱变育种在产油微生物生产DHA中的研究进展

doi:10.13560/j.cnki.biotech.bull.1985.2019-0637

摘要/Abstract

摘要： 二十二碳六烯酸（DHA）对维持人体正常的生理功能至关重要,具有极高的生物医学价值。产油微生物是指具有很强的脂肪酸合成能力,DHA占总脂肪酸含量相对较高的一类微生物,是获取DHA的新途径。诱变育种作为一种有效的变异手段在产油微生物选育与改良中显示了极为重要的作用和十分诱人的前景。综述了通过物理和化学诱变选育高产DHA菌株的机理及方法,其中物理诱变包括γ射线、紫外线、离子束和常压室温等离子体等;化学诱变包括甲基磺酸乙酯、亚硝基胍和硫酸二乙酯等;主要介绍了不同诱变育种方式在产油微生物生产DHA研究方面取得的成就,探讨了诱变育种的发展方向,提出了不同诱变育种方式相结合的复合育种的思路,以期对今后产油微生物生产DHA的研究有所启发。

关键词: DHA, 产油微生物, 诱变育种, 复合育种

Abstract: Docosahexaenoic acid（DHA）is essential in the maintenance of normal physiological function of human body and thus has significant biomedical value. Oleaginous microorganisms possess strong capacity to synthesize fatty acids and maintain relatively high percentage of DHA in total fatty acids. Mutagenesis is an effective and prospective breeding approach for the selection and improvement of oleaginous microorganisms. This review summarizes the methods and mechanisms of breeding the high-yield DHA strains by physical（γ-ray,ultraviolet,ion beam,and ARTP）or chemical（ethyl methanesulphonate,N-methyl-N’-nitro-N-nitrosoguanidine,and diethyl sulfate）agents. Also the paper introduces the research achievements on oleaginous microorganisms yielding DHA via varied mutation breeding methods,discusses the trends of mutation breeding,and put forward the idea of hybrid breeding by combining different breeding methods,its hoped that the study of DHA production by oleaginous microorganisms will be enlightened in the future.

Key words: DHA, oleaginous microorganisms, induced breeding, hybrid breeding

李建涛, 刘宪华, 何耀东, 汪光义. 诱变育种在产油微生物生产DHA中的研究进展[J]. 生物技术通报, 2020, 36(1): 110-115.

LI Jian-tao, LIU Xian-hua, HE Yao-dong, WANG Guang-yi. Mutagenesis Breeding in DHA Production by Oleaginous Microorganisms[J]. Biotechnology Bulletin, 2020, 36(1): 110-115.

参考文献

[1] Ghini V, Di Nunzio M, Tenori L, et al. Evidence of a DHA signature in the lipidome and metabolome of human hepatocytes[J]. Int J Mol Sci, 2017, 18(2):pii:E359.
[2] Vanu RR, Sijo JT, Shuo Y, et al.Effect of consuming novel foods consisting high oleic canola oil, barley β-glucan, and DHA on cardiovascular disease risk in humans:the CONFIDENCE(Canola Oil and Fibre with DHA Enhanced)study protocol for a randomized controlled trial[J]. Trials, 2015, 16(1):489.
[3] Zehr KR, et al.Omega-3 polyunsaturated fatty acids improve endot-helial function in humans at risk for atherosclerosis:A review[J]. Prostaglandins Other Lipid Mediat, 2017, 134:131-140.
[4] Deng DF, Ju ZY, Dominy WG, et al. Effect of replacing dietary menhaden oil with pollock or soybean oil on muscle fatty acid composition and growth performance of juvenile Pacific threadfin(Polydactylus sexfilis)[J]. Aquaculture, 2014, 422-423:91-97.
[5] 陈诚. 裂殖壶菌(Schizochytrium limacinum SR21)发酵制备二十二碳六烯酸(DHA)过程的初步研究[D]. 杭州:浙江大学, 2008.
[6] Wang Q, Sen B, Liu X, et al. Enhanced saturated fatty acids accu-mulation in cultures of newly-isolated strains of, Schizochytrium sp. and Thraustochytriidae sp. for large-scale biodiesel production[J]. Sci Total Environ, 2018, 631-632:994-1004.
[7] 王澍, 尹佳, 曹维, 等. 寇氏隐甲藻突变株发酵罐补糖发酵的研究[J]. 中国油脂, 2016, 41(1):41-44.
[8] Maria B, Bogdan R, Alexandru R, et al.Valorification of crude glycerol for pure fractions of docosahexaenoic acid and β-carotene production by using Schizochytrium limacinum and Blakeslea trispora[J]. Microbial Cell Factories, 2018, 17(1):97 .
[9] Zhang X, Zhang X, Li H, et al.Atmospheric and room temperature plasma(ARTP)as a new powerful mutagenesis tool[J]. Appl Microbiol Biotechnol, 2014, 98(12):5387-5396.
[10] Malook S, Qaisarani SA, Shabaz MK, et al.Mutation breeding approach to breed drought tolerant maize hybrids[J]. Int J Biosci, 2015, 6:427-436.
[11] Cheng R, Ma R, Li K, et al.Agrobacterium tumefaciens mediated transformation of marine microalgae Schizochytrium[J]. Microbiological Research, 2012, 167(3):179-186.
[12] Ren F, et al.Highly improved acarbose production of Actinomyces through the combination of ARTP and penicillin susceptible mutant screening[J]. World J Microbiol Biotechnol, 2017, 33(1):16.
[13] Yi Z, Xu M, Magnusdottir M, et al.Effect of physical, chemical mutagenesis and adaptive evolution on the marine diatom Phaeodactylum tricornutum for enhanced carotenoid accumulation[J]. Planta Medica, 2016, 82(S1):S1-S381.
[14] 黄鹭强, 王明兹, 李素萍, 等. 半导体激光对紫球藻生物学效应的影响[J]. 生物技术, 2002, 12(1):14-16.
[15] 佘隽, 田华, 陈涛, 等. 高产DHA寇氏隐甲藻突变株的筛选[J]. 食品科学, 2013, 34(17):230-235.
[16] 吕小义. 高产DHA裂壶藻突变株的选育与发酵工艺研究[D]. 武汉:武汉轻工大学, 2016.
[17] Morita R, et al.Heavy-ion beam mutagenesis identified an essential gene for chloroplast development under cold stress conditions during both early growth and tillering stages in rice[J]. Biosci Biotechnol Biochem, 2017, 81(2):271-282.
[18] Fu J, Chen T, Lu H, et al.Enhancement of docosahexaenoic acid production by low-energy ion implantation coupled with screening method based on Sudan black B staining in Schizochytrium sp.[J]. Bioresour Technol, 2016, 221:405-411.
[19] 王芝瑶, 马玉彬, 牟润芝, 等. 重离子诱变创制高产油微拟球藻新品种[J]. 生物工程学报, 2013(1):119-122.
[20] 蔡恺敏. Schizochytrium limacinum的诱变选育及发酵过程研究[D]. 杭州:浙江大学, 2008.
[21] 许永, 臧晓南, 徐涤, 等. 裂殖壶菌诱变筛选的研究[J]. 中国海洋大学学报:自然科学版, 2012, 42(12):54-58.
[22] 荣辉. 隐甲藻藻种的选育及其发酵条件的优化[D]. 海口:华南热带农业大学, 2007.
[23] 梁园梅, 刘瑛, 李晶晶, 等. 高产DHA破囊壶菌Aurantiochy-trium sp. PKU#SW7诱变株的筛选[J]. 微生物学通报, 2016, 43(2):457-464.
[24] Zhang X, Zhang XF, et al.Atmospheric and room temperature plasma(ARTP)as a new powerful mutagenesis tool[J]. Appl Microbiol Biotechnol, 2014, 98(12):5387-5396.
[25] Cheng G, et al.Breeding L-arginine-producing strains by a novel mutagenesis method:Atmospheric and room temperature plasma(ARTP)[J]. Prep Biochem Biotechnol, 2016, 5:509-516.
[26] Qian QZ, Xiao FZ, Li YW, et al.Quantitative evaluation of DNA damage and mutation rate by atmospheric and room-temperature plasma(ARTP)and conventional mutagenesis[J]. Appl Microbiol Biotechnol, 2015, 99(13):5639-5646.
[27] Li HP, Wang ZB, Nan G, et al.Studies on the physical characteri-stics of the radio-frequency atmospheric-pressure glow discharge plasmas for the genome mutation of Methylosinus trichosporium[J]. IEEE Trans Plasma Sci, 2012, 40(11):2853-2860.
[28] 龚定芳, 熊祎楠, 孙佼文. 常压室温等离子体诱变选育DHA高产菌株[J]. 中国油脂, 2019, 44(3):118-122.
[29] Zhao B, et al.Enhancement of Schizochytrium DHA synthesis by plasma mutagenesis aided with malonic acid and zeocin screening[J]. Appl Microbiol Biotechnol, 2018, 102(5):2351-2361.
[30] 袁军, 赵犇, 孙梦玉. 常压室温等离子体(ARTP)诱变快速选育高产DHA的裂殖壶菌突变株[J]. 生物技术通报, 2015, 31(10):199-204.
[31] Fang M, Jin L, Zhang C, et al.Rapid mutation of Spirulina platensis by a new mutagenesis system of atmospheric and room temperature plasmas(ARTP)and generation of a mutant library with diverse phenotypes[J]. PLoS One, 2013, 8(10):e77046.
[32] Liu J, Pei G, Diao J, et al.Screening and transcriptomic analysis of Crypthecodinium cohnii mutants with high growth and lipid content using the acetyl-CoA carboxylase inhibitor sethoxydim[J]. Appl Microbiol Biotechnol, 2017, 101(15):6179-6191.
[33] Ahmad Khan S, Rahman LU, Verma R, et al.Physical and chemical mutagenesis in Stevia rebaudiana:variant generation with higher UGT expression and glycosidic profile but with low photosynthetic capabilities[J]. Acta Physiologiae Plantarum, 2016, 38(1):4.
[34] 赵爱娟. 海洋微藻的诱变育种及其脂肪酸的测定[D]. 南京:南京理工大学, 2005.
[35] 陈浩. 裂殖壶菌培养基优化、菌种选育及遗传转化体系的研究[D]. 青岛:中国海洋大学, 2014.
[36] 陈金卿, 陈俊煌, 吴美琼, 等. 一种DHA产生菌双鞭甲藻的诱变筛选方法:中国, CN 102391955A[P].2012-03-28.
[37] 李慧玲, 刘永梅. 诱变选育高产DHA裂殖壶菌突变株[J]. 食品科技, 2015, 40(9):12-16.
[38] Lian M, Huang H, Ren L, et al.Increase of docosahexaenoic acid Production by Schizochytrium sp. through mutagenesis and enzyme assay[J]. Appl Biochem Biotechnol, 2010, 162(4):935-941.
[39] 王申强. 裂殖壶菌产DHA的发酵工艺研究及高产菌株选育[D]. 无锡:江南大学, 2013.
[40] 赵犇, 王武, 李昌灵. EMS-ARTP复合诱变选育高产DHA裂殖壶菌[J]. 食品与机械, 2018, 196(2):25-30.
[41] 石婷婷. 产油脂微生物菌株的选育[D]. 济南:山东轻工业学院, 2012.
[42] 王婷婷, 詹晓北, 郑志永, 等. 高产花生四烯酸高山被孢霉菌株的诱变筛选[J]. 工业微生物, 2013, 43(4):44-50.