CN113801798B - 食腺嘌呤节孢酵母a50菌株、所产胞外多糖及其应用 - Google Patents
食腺嘌呤节孢酵母a50菌株、所产胞外多糖及其应用 Download PDFInfo
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- CN113801798B CN113801798B CN202110881679.XA CN202110881679A CN113801798B CN 113801798 B CN113801798 B CN 113801798B CN 202110881679 A CN202110881679 A CN 202110881679A CN 113801798 B CN113801798 B CN 113801798B
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- extracellular polysaccharide
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Abstract
本发明属于微生物技术领域，具体涉及一种Blastobotrys adeninivorans A50菌株、及其产生的胞外多糖，并公开了胞外多糖在结合胆汁酸中的作用。本发明的菌株分离自渥堆过程的六堡茶样品，通过分子学和形态学鉴定为Blastobotrys adeninivorans菌。该菌株的胞外多糖含有78.08％的总糖、17.81％的糖醛酸，紫外光谱显示不含核酸蛋白、红外光谱表明该胞外多糖是酸性多糖，且是一种硫酸多糖，同时存在α‑吡喃环和呋喃环。该胞外多糖在体外具有较好的结合胆汁酸作用，半抑制浓度IC50值为64.325mg/ml，显示出具有潜在降血脂的能力，在高脂血症的防治和治疗等方面具有良好应用前景。
Description
技术领域
本发明属于微生物技术领域，更具体的，涉及一株可产胞外多糖具有胆汁酸结合作用的Blastobotrys adeninivorans A50菌株及其应用。
背景技术
高脂血症是指血浆(或血清)中一种或者多种脂质成分异常增高为特征的病症。该症引起的动脉粥样硬化是心血管疾病发生的重要因素。在肝中被转换成为胆汁酸是胆固醇在体内代谢的主要去路。肝脏生成的胆汁酸95％以上会被人体重新吸收，余下未被吸收的小部分最终会随着粪便排出。胆汁酸结合剂可在小肠与胆汁酸结合形成不溶性复合物并排出体外，导致肝脏或者血液中的胆固醇加快转变为胆汁酸，从而发挥降低体内胆固醇的作用。
胆汁酸结合剂有两大类：阴离子交换树脂类和膳食纤维多糖类。服用一些膳食纤维多糖如燕麦和大麦的β-葡聚糖，水果中的果胶，瓜尔胶会导致粪便中胆汁酸的***量增加至35％-65％。微生物多糖因其成本低、易生产，不受季节的影响而在食品、制药等各个领域中被广泛利用。然而相比植物多糖，对于微生物胞外多糖的胆汁酸结合活性研究较少，这不利微生物资源的开发。
六堡茶被认为是健康、养生的休闲饮品，已有研究六堡茶多糖能够结合血液中的胆汁酸。特殊的制茶工艺使六堡茶独特品质的形成离不开渥堆发酵过程中微生物的作用。已有研究报道冠突散囊菌的胞外多糖是茯砖茶多糖活性成分之一，具有降脂活性。然而，关于六堡茶中的Blastobotrys adeninivorans胞外多糖的功能信息尚未见报道。
发明内容
为克服现有技术中的不足，本发明提供如下技术方案：
本发明提供了一株可产胞外多糖具有胆汁酸结合作用的Blastobotrysadeninivorans A50菌株，所述Blastobotrys adeninivorans A50于2021年04月29日保藏于中国典型培养物保藏中心(CCTCC)，保藏编号为CCTCC NO:M 2021485。
上述Blastobotrys adeninivorans菌株属于子囊菌门，酵母纲，酵母目，Trichomonascaceae科，Blastobotrys属。
(1)本发明的A50菌株是从广西壮族自治区梧州市六堡茶企业渥堆过程中发酵茶叶获得，鉴定结果显示，所述食腺嘌呤节孢酵母A50在PDA培养基上菌落呈白色、干燥，形态为圆形、不透明；显微镜下可观察到细胞呈圆形或类圆形。
进一步地，所述A50菌株在PDB培养基中产胞外多糖。该菌的发酵液经过离心收集上清液、再加入3倍体积无水乙醇沉淀两次，晾干。加入蒸馏水溶解，用Sevag法除蛋白，再加入三倍体积95％的乙醇，离心沉淀。沉淀加适量蒸馏水溶解后装入截留分子量为8000Da-12000 Da的透析袋，随后将糖溶液进行冷冻干燥，命名为EPS-A50。
EPS-A50的组成为78.08％的总糖、17.81％的糖醛酸。紫外光谱在260nm-280nm没有吸收峰，表明不含核酸或蛋白。红外光谱表明该胞外多糖是酸性多糖且是一种硫酸多糖，；单糖组成有甘露糖，同时存在α-吡喃环和呋喃环。EPS-A50的分子量大小为320.799KDa(26.28％)，26.618KDa(72.30％)，1.786KDa(1.42％)，单糖组成由Man、GalA和Gal组成，摩尔比分别是0.99：1.34：39.02。
更进一步的，该胞外多糖在浓度为10-110mg/mL时具有23.81％-79.24％的体外结合胆汁酸作用，半抑制浓度IC50值为64.325mg/ml。因此，可应用于制备成微生物菌剂，例如，可以作为胆汁酸结合剂，用作降血脂，尤其是可制备预防和/或治疗高脂血症中使用。
此微生物菌剂可以制备成食品、保健品或药品中可接受的制剂。由于该菌株来自六堡茶，因而成分安全可靠，应用性强。
与现有技术相比，本发明具有以下优点：菌株来源可靠，成分安全。采用微生物发酵技术，易于实现扩大生产。菌株所产胞外多糖的结合胆汁酸作用效果好，应用性强。
附图说明
图1为A50菌株的显微镜图。
图2为A50菌株Neighbour-Joining算法***进化树图。
图3为EPS-A50的紫外光谱图。
图4为EPS-A50的红外光谱图。
图5为EPS-A50的分子量大小图。
图6为EPS-A50的单糖组成图。
图7为EPS-A50体外结合胆汁酸的效果图。
具体实施方式
下面将结合本发明实施例，对本发明的技术方案进行清楚、完整地描述，显然，所描述的实施例仅仅是本发明一部分实施例，而不是全部的实施例。基于本发明中的实施例，本领域普通技术人员在没有做出创造性劳动前提下所获得的所有其他实施例，都属于本发明保护的范围。实施例中未注明具体条件者，按照常规条件或制造商建议的条件进行。所用试剂或仪器未注明生产厂商者，均为可以通过市售购买获得的常规产品。
实施例1 Blastobotrys adeninivorans A50菌株的分离、培养与鉴定
1、菌株分离与培养
(1)收集菌落：取新鲜的处于渥堆过程中的六堡茶样品(采集地：广西壮族自治区梧州市六堡茶企业渥堆过程中发酵茶叶样品)2g，用剪刀剪碎后倒入装有20mL无菌水的蓝口瓶，80r/min条件下振荡30min，使附着在茶叶上的微生物脱落。
(2)菌落纯化：取1mL菌落原液，分别稀释成10-1、10-2、10-3、10-4、10-5、10-6的梯度浓度菌悬液，吸取200μL的菌悬液，打入PDA中，用一次性涂布棒涂布均匀。放置28℃的培养箱培养2到3天。待涂布培养皿长出菌落后，通过肉眼观察，分类，挑取形态不同的菌株的菌落边缘接种到PDA，重复纯化4次以上，直至成为单菌落。
2、菌株形态学鉴定
(1)A50菌株在PDA上菌落呈白色、干燥，形态为圆形、不透明。
(2)显微镜观察：如图1所示，细胞呈圆形或类圆形。
3、分子鉴定
(1)首先进行16S rRNA测序：将纯化后的单菌落接种于PDA，在28℃的培养箱培养12到24小时。取50μL灭菌过的树脂溶液于八连管，用牙签挑取极少部分的菌体放置于八连管中研磨1min。八连管在金属浴100℃下保持10min，离心取上清液即为模板DNA。对模板DNA片段进行扩增，送到生物公司进行测序。获得A50菌株的16S rRNA的序列，如SEQ ID NO.1所示。
(2)在EzTaxon数据库(http://www.ezbiocloud.net/eztaxon)中收集所有食腺嘌呤节孢酵母属的菌种16S rRNA的序列。所有的序列采用软件CLUSTAL_X进行对齐和剪切，采用软件MEGA X构建进化树，算法为Neighbour-joining和Maximum-likelihood算法(Bootstrap：1000replications)。
利用序列比对进行***发育分析，进化树结果见图2。结果显示，该A50菌株属于Blastobotrys，并且最相近的菌种是Blastobotrys adeninivorans，并将其进行保藏，A50菌株已于2021年04月29日保藏于中国典型培养物保藏中心(CCTCC)，保藏编号为CCTCC NO:M 2021485，保藏地址：中国武汉武汉大学。
实施例2 Blastobotrys adeninivoransA50菌株的发酵培养
1、种子液培养
菌株按照编号从冻存管盒里取出，放置在室温下溶解，接种到PDA，在28℃下培养24小时，然后用接种挑取单菌落接入YEPD液体培养基(蛋白胨20.0g，酵母粉10.0g，葡萄糖20.0g，蒸馏水1000.0mL)，在28℃，120r/min的条件下发酵18-24小时，调节菌液在600nm处的吸光值为1.0左右，即为种子液。
2、扩大培养
按照5％的接种比例接到含有250mL PDB的500mL锥形瓶，在28℃，120r/min的条件下发酵168小时。
实施例3 Blastobotrys adeninivoransA50菌株所产胞外多糖EPS-A50的提取、纯化及组份测定
1、提取、纯化
发酵结束后，发酵液在6000r/min，20min的条件下离心，加入三倍体积无水乙醇，4℃冰箱过夜沉淀，沉淀晾干后加入一定体积的蒸馏水重融，再加入3倍体积的95％乙醇，4℃过夜。收集沉淀，晾干。加入蒸馏水溶解配制成为1％的糖溶液，用Sevag法除蛋白。将正丁醇与蒸馏水等体积混合，超声10min，取上层溶液为水饱和的正丁醇溶液。在上述正丁醇溶液加入4倍体积氯仿得到Sevag试剂。在糖溶液中加入三分之一倍体积的Sevag试剂，磁力搅拌器搅拌20min，倒入分液漏斗静止直至看到明显的分层，将下层的有机试剂和中层的变形蛋白除去。收集上层为除蛋白后的糖溶液。重复除蛋白操作，直至看不到乳白色的沉淀。随后在60℃条件下旋转蒸发除去有机试剂。加入三倍体积95％的乙醇并在4℃冰箱过夜，离心收集沉淀。沉淀加适量蒸馏水溶解后装入截留分子量为8000Da-12000 Da的透析袋，在100倍体积蒸馏水中透析，每隔4小时换一次水，共透析48小时。透析后，将糖溶液进行冷冻干燥，得到EPS-A50。
2、组份测定
总糖、糖醛酸和蛋白质含量的测定方法参照文献略有修改。结果表明，EPS-A50含有78.08±1.59％的总糖、17.81±1.21％的糖醛酸，不含蛋白质。
实施例4 Blastobotrys adeninivorans A50菌株所产胞外多糖EPS-A50的紫外、红外光谱分析
1、紫外光谱
将胞外多糖样品溶于蒸馏水并调节到合适的浓度，随后用紫外光谱仪(日本岛津公司，UV 2501PC)进行扫描分析，扫描波长在200nm-400nm范围。实验结果如图3所示，EPS-A50在260nm-280nm处没有吸收峰，表明不含有核酸和蛋白质。
2、红外光谱
称取胞外多糖样品3mg和溴化钾固体粉末270mg，混合研磨至样品颗粒没有晶状体后用压片机制成均匀的薄片。用傅里叶变换红外光谱仪(Fourier transform-infrared，FT-IR)进行扫描，扫描范围为4000cm-1-400cm-1。
如图4所示EPS-A50在3407.47cm-1处出现强烈吸收峰可归因于O-H键的伸缩振动峰，表明存在羟基；2933.45cm-1处的峰是糖链上的-CH2或-CH的C-H键的伸缩振动峰。以上是多糖特征吸收峰。1639.50cm-1处附近的峰归因于束缚水的存在。1418.50cm-1处出峰可能是羧基或羧酸盐的特征，表明胞外多糖都是酸性多糖，这与理化指标的测定一致。多糖在1239.15cm-1处有吸收峰，这是S＝O的伸缩振动峰，表明是一种硫酸多糖。1100cm-1-1010cm-1有3个吸收峰则代表有吡喃环，而有2个吸收峰则代表有呋喃环。结果表明多糖只有2个峰，表明含呋喃环。EPS-A50在1079.18cm-1处是吡喃环的醚键C-O-C和羟基的吸收峰。多糖在1021.13cm-1处是C-O伸缩振动峰。931.67cm-1处出峰表明可能含有甘露糖。在890cm-1和812cm-1没有吸收峰(β构型的特定信号)，而在850cm-1左右出峰，表明含有α-糖苷键。EPS-A50在765.13cm-1有吸收峰是由α-吡喃环对称伸缩振动引起的。
以上结果说明：胞外多糖都是酸性多糖且是一种硫酸多糖，单糖组成有甘露糖。EPS-A50同时存在α-吡喃环和呋喃环。
实施例5 Blastobotrys adeninivorans A50所产胞外多糖EPS-A50的分子量及单糖组份检测
1、分子量测定
采用凝胶渗透色谱(Gel Permeation Chromatography，GPC)法测定胞外多糖的分子量。胞外多糖和系列葡聚糖标品(580、29330、10730、31420、63350、143400、280500、841700、2560000和7500000Da)溶于超纯水，配制成1mg/mL的溶液。色谱条件：Waters GPC1515型，配备示差折光检测器，色谱柱：WATERS Ultrahydrogel 500column(7.8×300mm)、WATERS Ultrahydrogel 250column(7.8×300mm)和WATERS Ultrahydrogel 120column(7.8×300mm)串联合用。流动相：超纯水；流速：1.0mL/min；柱温：30℃；进样量：40μL。
结果如图5所示，图5(a)是葡聚糖标品的标准曲线，R2＝0.9985说明线性范围较好，方法可靠。图5(b)是EPS-A50的GPC图，把保留时间带入标准曲线，可知EPS-A50的分子量大小为320.799KDa(26.28％)，26.618KDa(72.30％)，1.786KDa(1.42％)。
2、单糖组成测定
多糖的水解：取胞外多糖粉末0.0500g置于水解管中，加1mL的HCl(3.0mol/L)，封口，混匀，110℃水解4-6小时，放冷，用3.0mol/L的NaOH调节pH值至中性，吸取400μL进行衍生化处理。
单标和梯度混标的配制。(1)单标。各单糖对照品(***糖、甘露糖、葡萄糖、半乳糖、半乳糖醛酸、葡萄糖醛酸、岩藻糖和鼠李糖)溶于超纯水，浓度为2mmol/L，。(2)混标。单糖对照品共溶于超纯水，使得每个标品的浓度为1.6mg/mL，后稀释为1.6、1.2、1.0、0.8、0.4、0.2mg/mL共6个浓度的梯度混标。
单糖衍生：取标品溶液(单标和梯度混标)及水解好的多糖样品400μL，加0.5mol/L的PMP-甲醇溶液与0.3mol/L的NaOH各400μL，混匀，70℃水浴反应100min。再加入500μL的0.3mol/L的HCl，用三氯甲烷萃取PMP试剂3次，每次5mL，弃去三氯甲烷层，水层离心后，取上清液1mL过0.45μm有机系滤膜，进行高效液相色谱(High Performance LiquidChromatography，HPLC)分析。
采用Waters 2984LC型高效液相色谱仪进行分析。色谱条件：Waters C18色谱柱(4.6×150mm，5μm)；以乙腈-0.02mol/L的乙酸铵溶液(17：83)为流动相色谱条件；柱温30℃；流速1.0mL/min；检测波长250nm，进样量10μL。
由图6(a)和图6(b)所示，PMP柱前衍生法可有效将8种单糖标品分开，把保留时间及峰面积带入标准曲线计算EPS-A50的单糖组成，可知EPS-A50由Man、GalA和Gal组成，摩尔比分别是0.99：1.34：39.02。
实施例6食腺嘌呤节孢酵母所产胞外多糖EPS-A50体外结合胆汁酸能力。
首先，配制胆汁酸储备溶液：甘氨胆酸(9mmol/L)、鹅去氧胆酸(9mmol/L)、甘氨脱氧胆酸(9mmol/L)，牛磺鹅脱氧胆酸(4.5mmol/L)，牛磺脱氧胆酸(4.5mmol/L)溶于pH＝6.8的磷酸盐缓冲溶液，即配制成为36mmol/L的胆汁酸储备溶液。将储备溶液储存在-20℃的冰箱，并在使用前稀释为0.72μmol/mL的胆汁酸工作液。
参考Wu等的方案并略作修改。将10mg/mL-110mg/mL的胞外多糖，装入15×150mm的玻璃试管，加入1mL，0.01mol/L的HCl，于37℃，120r/min模拟胃的酸性消化2小时。用0.2mol/L的NaOH将样品pH调节至7-7.5。向每个待测样品中加入4mL胆汁酸工作液(0.72μmol/mL)和5mL胰酶(10mg/mL，溶于pH＝6.8的磷酸缓冲液)，于37℃，120r/min条件下模拟体外肠消化2小时。混合物在6000r/min的条件下离心20min，取上清液存于-20℃待测。以蒸馏水作为阴性对照。
根据总胆汁酸试剂盒的测定方法对上清液中胆汁酸含量进行检测。胆汁酸含量的公式为：
其中，酶标仪(Infinite M200 PRO)读取405nm处吸光度A0，3min后再读取吸光度A1，ΔA＝A1-A0。ΔA测定是样品的ΔA值，ΔA校准是校准品的ΔA值，胆汁酸校准品的浓度为50μmol/L。
结果如图7所示，EPS-A50半抑制浓度IC50值为64.325mg/ml。表明EPS-A50有潜在降血脂的能力，有望成为高血脂症的预防和治疗的药物，而该菌株有望开发成为商业菌株。
以上对本发明的实施方式作了详细说明，但本发明不限于所描述的实施方式。对于本领域的技术人员而言，在不脱离本发明原理和精神的情况下，对这些实施方式进行多种变化、修改、替换和变型，仍落入本发明的保护范围内。
序列表
<110> 广西大学
<120> 食腺嘌呤节孢酵母 A50菌株、所产胞外多糖及其应用
<130> ZM211185CS
<160> 1
<170> SIPOSequenceListing 1.0
<210> 1
<211> 500
<212> DNA
<213> 2 Ambystoma laterale x Ambystoma jeffersonianum
<400> 1
gtcttcggaa tttttatcct taacacctgt gaactattgt aattttttgc tttggtcctt 60
cggggccaga ggatttaatc aaaacctttt attattaacc tgagtctgaa aattgaaaaa 120
ttgaattatt caaaactttc aacaacggat ctcttggttc tcgcatcgat gaagaacgca 180
gcgaactgcg ataagtaatg tgaattgcag aattttgtga atcatcgaat ctttgaacgc 240
acattgcgcc ttttggtatt ccagaaggca tacctgtttg agagtcattt cttcctcaat 300
cctcggattg gtgttgactc gagcgttgcg aaacgctgag tcgaaaagaa ttggcaggca 360
ggaattcagt gctttacagt gtcttaggtt ttaccaacta cgctggccta gtaactgttt 420
tctgagctgg cctttataac agttctttta aaagtttgac ctcaaatcag gcaagactac 480
ccgctgaact taagcatatc 500
Claims (8)
1.食腺嘌呤节孢酵母（Blastobotrys adeninivorans）A50菌株在制备胆汁酸结合剂中的应用，其特征在于，所述Blastobotrys adeninivorans A50菌株于2021年04月29日保藏于中国典型培养物保藏中心(CCTCC)，保藏编号为CCTCC NO: M 2021485。
2.根据权利要求1所述应用，其特征在于，所述胆汁酸结合剂应用于食品或药品中。
3.根据权利要求1所述应用，其特征在于，将食腺嘌呤节孢酵母Blastobotrys adeninivorans A50菌株常规发酵，取上清液。
4.一种胞外多糖EPS-A50，其特征在于，所述胞外多糖由食腺嘌呤节孢酵母Blastobotrys adeninivorans A50菌株按照常规发酵，取上清液后得到；所述Blastobotrys adeninivorans A50菌株于2021年04月29日保藏于中国典型培养物保藏中心(CCTCC)，保藏编号为CCTCC NO: M 2021485。
5.权利要求4所述胞外多糖EPS-A50在制备预防和/或治疗降血脂的药物中的应用。
6.根据权利要求5所述应用，其特征在于，是在制备预防和/或治疗高脂血症的药物中的应用。
7.一株食腺嘌呤节孢酵母（Blastobotrys adeninivorans）A50菌株，其特征在于，所述Blastobotrys adeninivorans A50菌株于2021年04月29日保藏于中国典型培养物保藏中心(CCTCC)，保藏编号为CCTCC NO: M 2021485。
8.一种微生物菌剂，其特征在于，包括权利要求7所述食腺嘌呤节孢酵母Blastobotrys adeninivorans A50菌株。
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Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101709093A (zh) * | 2009-12-17 | 2010-05-19 | 广西大学 | 一种滇桂艾纳香水溶性多糖的制备方法 |
| CN105779297A (zh) * | 2014-12-16 | 2016-07-20 | 勐海茶业有限责任公司 | 一株产高活性多酚氧化酶的酵母菌及其在普洱茶生产中的应用 |
| CN109055247A (zh) * | 2018-07-13 | 2018-12-21 | 云南中茶茶业有限公司 | 一株酵母菌株及其加工熟普茶的方法 |
| CN109385485A (zh) * | 2017-08-09 | 2019-02-26 | 勐海茶业有限责任公司 | Dna条形码、引物、试剂盒、方法和应用 |
| RU205304U1 (ru) * | 2021-04-12 | 2021-07-08 | Федеральное государственное бюджетное образовательное учреждение высшего образования "Тульский государственный университет" (ТулГУ) | Устройство для экспресс-анализа загрязненности воды биоразлогаемыми органическими соединениями |
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Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101709093A (zh) * | 2009-12-17 | 2010-05-19 | 广西大学 | 一种滇桂艾纳香水溶性多糖的制备方法 |
| CN105779297A (zh) * | 2014-12-16 | 2016-07-20 | 勐海茶业有限责任公司 | 一株产高活性多酚氧化酶的酵母菌及其在普洱茶生产中的应用 |
| CN109385485A (zh) * | 2017-08-09 | 2019-02-26 | 勐海茶业有限责任公司 | Dna条形码、引物、试剂盒、方法和应用 |
| CN109055247A (zh) * | 2018-07-13 | 2018-12-21 | 云南中茶茶业有限公司 | 一株酵母菌株及其加工熟普茶的方法 |
| RU205304U1 (ru) * | 2021-04-12 | 2021-07-08 | Федеральное государственное бюджетное образовательное учреждение высшего образования "Тульский государственный университет" (ТулГУ) | Устройство для экспресс-анализа загрязненности воды биоразлогаемыми органическими соединениями |
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