地毯草黄单胞菌分泌黄原胶的电镜观察
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摘要
为了研究地毯草黄单胞菌(Xanthomonas axonopodis)FJAT-10151产黄原胶的分泌行为,采用比浊法测定地毯草黄单胞菌的生长曲线,干重法测定黄原胶产量,2,4-二硝基苯肼显色法测定黄原胶中丙酮酸含量变化,通过扫描电子显微镜镜(SEM)和透射电子显微镜镜(TEM)观察地毯草黄单胞菌产黄原胶的分泌行为。FJAT-10151在发酵时间0～26 h为对数期,26～72 h为稳定期。菌株FJAT-10151为短杆状(约2μm),端生鞭毛(约7.9μm),菌体外面形成了一层较厚的荚膜,且许多菌体通过黄原胶连在一起,分泌的黄原胶呈棒状和网状2种形态,聚集或缠绕在细胞表面,其含量随发酵时间的延长而增加。菌株FJAT-10151发酵72 h可获得高品质黄原胶,其产量及丙酮酸含量最高。
In order to study the xanthan gum secreted by Xanthomonas axonopodis FJAT-10151, The growth curve of FJAT-10151 was determined by the turbidimetric method. The yield of xanthan gum was determined by the dry-weighing method. The content of pyruvic acid was determined by the 2,4-dinitrobenzene hydrazine chromogenic method. The secretion of xanthan gum from X. axonopodis was studied by SEM and TEM. The result showed that the logarithmic phase and stable phase of FJAT-10151 was 0–26 h and 26–72 h, respectively. FJAT-10151 was a short rod of 2 μm long with 7.9 μm of flagellum on the ends, and formed a thick capsule outside, and many bacteria together with xanthan gum. Xanthan gum had flake and filamentous forms, and gathered or wrapped around FJAT-10151 surface. The yield of xanthan gum increased with the extension of fermentation time. The yield of xanthan gum and pyruvic acid reached the highest at the fermentation time of 72 h. We could obtain high quality of xanthan gum at the fermentation time of 72 h.
In order to study the xanthan gum secreted by Xanthomonas axonopodis FJAT-10151, The growth curve of FJAT-10151 was determined by the turbidimetric method. The yield of xanthan gum was determined by the dry-weighing method. The content of pyruvic acid was determined by the 2,4-dinitrobenzene hydrazine chromogenic method. The secretion of xanthan gum from X. axonopodis was studied by SEM and TEM. The result showed that the logarithmic phase and stable phase of FJAT-10151 was 0–26 h and 26–72 h, respectively. FJAT-10151 was a short rod of 2 μm long with 7.9 μm of flagellum on the ends, and formed a thick capsule outside, and many bacteria together with xanthan gum. Xanthan gum had flake and filamentous forms, and gathered or wrapped around FJAT-10151 surface. The yield of xanthan gum increased with the extension of fermentation time. The yield of xanthan gum and pyruvic acid reached the highest at the fermentation time of 72 h. We could obtain high quality of xanthan gum at the fermentation time of 72 h.
引文
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[2]李淑彬,方茂,林晓云,等.地毯草黄单胞菌ABB-2对红掌防御相关生化指标的影响[J].华南师范大学学报(自然科学版),2013,45(1):79-84.
[3]Osdaghi E,Zademohamad A A.Phaseolus lunatus,a new host of Xanthomonas axonopodis pv.phaseoli in Iran[J].Journal of Phytopathology,2016,164(1):56-60.
[4]Khodakaramian G,Swings J.AFLP fingerprinting of the strains of Xanthomonas axonopodisinducing citrus canker disease in southern Iran[J].Journal of Phytopathology,2002,150(4-5):227-231.
[5]Pegos V R,Canevarolo R R,Sampaio A P,et al.Xanthan gum removal for 1H-NMR analysis of the intracellular metabolome of the bacteria Xanthomonas axonopodis pv.citri306[J].Metabolites,2014,4(2):218-231.
[6]Rojas R,Nishidomi S,Nepomuceno R,et al.Glutamate transport and xanthan gum production in the plant pathogen Xanthomonas axonopodis pv.citri[J].World Journal of Microbiology&Biotechnology,2013,29(11):2173-2180.
[7]郑梅霞,朱育菁,刘波,等.微生物多糖胶质高产菌株的筛选与鉴定[J].食品科学,2016,37(15):171-178.
[8]García-Ochoa F,Santos V E,Casas J A,et al.Xanthan gum:production,recovery,and properties[J].Biotechnology Advances,2000,18(7):549-579.
[9]欧杰,李柏林.甘蓝黑腐病黄单胞菌胞膜离子通道透性对黄原胶生物合成的影响[J].食品科学,2003,24(1):43-48.
[10]秦志刚,詹晓北,贾薇,等.高丙酮酸黄原胶发酵与流变特性研究[J].食品与发酵工业,2007,33(1):32-35.
[11]刘如林,李志明,施贻君,等.黄原胶中丙酮酸含量的研究[J].微生物学杂志,1989,9(4):1-6.
[12]杨建华,孙丹,杨茂,等.利福平及杆菌肽抗性突变对黄单胞菌产胶率的影响[J].实验室科学,2006(6):52-54.
[13]牛淑敏,于松涛,王玉香,等.野油菜黄单胞菌8004胞外酶及多糖合成正负调控基因在X.c.NK01菌中功能初探[J].南开大学学报(自然科学版),1997,1:76-81,97.
[14]李元鑫,尹丽华,亓烨.一株产高黏度耐酸性黄原胶生产菌的选育及其发酵工艺研究[J].食品科学,2012(9):211-216.
[15]赵大健,王锐.黄原胶及其在食品工业上的应用[J].食品与发酵工业,1986,3:51-57.
[16]李鸿雁,陈庆森,阎亚丽,等.培养基组成对冰核生产菌Xanthomonas ampelina TS206产胶率的影响[J].食品科学,2006,27(2):116-119.
[17]刘元涛,董学前,王伟,等.鞘氨醇单胞菌发酵生产韦兰胶培养基优化研究[J].中国酿造,2016,35(1):73-77.
[18]董学前,刘元涛,王伟,等.鞘氨醇单胞菌发酵生产韦兰胶的研究[J].发酵科技通讯,2015,44(4):30-33.
[19]陈梅春,王阶平,肖荣凤,等.地衣芽胞杆菌FJAT-4脂肽结构鉴定及其对尖孢镰刀菌的抑制作用[J].微生物学报,2017,57(12):1924-1934.
[20]陆月良,颜永碧.一种保存细菌鞭毛的电镜负染色方法[J].第二军医大学学报,1994,15(1):28.
[21]沈萍,陈向东.微生物学[M].北京:高等教育出版社,2006:135-136.
[22]朱艳蕾.细菌生长曲线测定实验方法的研究[J].微生物学杂志,2016,36(5):108-112.
[23]李晓颖.茄科青枯菌致病力丧失相关基因的功能研究[D].广州:华南农业大学,2016.
[24]Hsu C H,Lo Y M.Characterization of xanthan gum biosynthesis in a centrifugal,packed-bed reactor using metabolic flux analysis[J].Process Biochemistry,2003,38(11):1617-1625.
[25]赵润宝,詹晓北,郑志永,等.添加游离丙酮酸对黄单胞杆菌产黄原胶中丙酮酸功能基团含量的影响[J].食品研究与开发,2006,27(1):14-17.
[26]莫晓燕,吉丽娜,詹谷宇.黄原胶发酵培养基优化研究[J].工业微生物,2003,33(2):15-18.
[27]张建国,黄勋娟.一株野油菜黄单胞菌产黄原胶的性质[J].食品科学,2014,35(23):29-32.
[28]李艳青,智丽丽,陈惠敏.黄原胶分子形貌的原子力显微镜研究[C]//新疆物理学会第六次会员代表大会暨学术交流会论文集,2010:23-26.
[29]周盛华,黄龙,张洪斌.黄原胶结构、性能及其应用的研究[J].食品科技,2008,33(7):156-160.
[30]何飞燕,林华.黄原胶结构、性能以及在食品加工中的应用[J].广西职业技术学院学报,2009,2(5):1-4.
[31]王超.水稻黄单胞菌水稻致病变种的xrv A基因的功能鉴定[D].南宁:广西大学,2006.
[32]钱韦.两种植物病原黄单胞菌基因组中同义密码子使用的分析[J].植物病理学报,2004,34(2):97-106.
[33]陆光涛,唐纪良,危广宁,等.野油菜黄单胞菌野油菜致病变种8004菌株wxcA基因与EPS的产量有关[J].生物工程学报,2004,20(4):477-483.