CN105670968B - 一种海洋天然抗胶质瘤活性物质及制备和用途 - Google Patents
一种海洋天然抗胶质瘤活性物质及制备和用途 Download PDFInfo
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Abstract
一种海洋天然抗胶质瘤活性物质及制备和用途，本发明提供一株放线菌菲律宾链霉菌，分类命名为Streptomyces filipinensis ZQ‑22。是从红树林下土壤中分离培养获得。利用本发明放线菌菲律宾链霉菌，经发酵、提取分离纯化制备得到具有抗胶质瘤活性的化合物Bagremycin C。该化合物显著抑制不同胶质瘤细胞的增殖，并显著诱导肿瘤细胞凋亡和影响肿瘤细胞周期，可在制备治疗胶质瘤药物中的应用。Bagremycin C化学结构式为：
Description
技术领域
本发明属于海洋生物技术领域，涉及从海洋放线菌菲律宾链霉菌(Streptomycesfilipinensis ZQ-22)中制备一种新的海洋天然抗胶质瘤活性物质Bagremycin C的方法以及该化合物在制备抗胶质瘤药物方面的应用。
背景技术
胶质瘤是中枢神经系统最常见和死亡率极高的脑肿瘤，约占所有恶性脑肿瘤的70％。手术后辅以放疗和化疗相结合是目前胶质瘤最常用的治疗方法。由于胶质瘤累及许多重要脑功能区，使得胶质瘤切除手术极为困难，所以，与其它肿瘤相比，药物对胶质瘤的治疗就更为重要。但是，目前全球用于治疗胶质瘤的药物严重缺乏，只有包括替莫唑胺(TMZ)、卡氮芥(BCNU)和环己亚硝脲(CCNU)等为数不多的几种。现有这些抗胶质瘤药大多都是以细胞毒为治疗原理的烷化剂对脑胶质瘤的治疗存在严重不足，主要包括：(1)毒副作用大，对患者产生较大的不良反应；(2)胶质瘤细胞对药物耐药性的不断增加严重影响药物的疗效；(3)血脑屏障的阻碍限制了治疗脑胶质瘤药物到达脑内发挥更有效的疗效。很显然，临床上迫切需要疗效好、毒副作用小、容易透过血脑屏障、作用机制独特的治疗胶质瘤新型药物。
源自天然产物的药物为人类的健康贡献巨大，特别是抗感染药物和抗肿瘤药物。据统计，有超过大约50％的抗肿瘤药物与天然产物有关。但是，遗憾的是目前全球还没有与天然产物有关的抗胶质瘤药物在临床上使用。特殊的生态环境使海洋微生物形成了独特的有别于陆生微生物的新陈代谢途径和适应机制，从而产生众多结构新颖和生物活性多样的代谢产物，是发现抗胶质瘤新药的重要新资源。
发明内容
本发明的目的是提供一株放线菌菲律宾链霉菌(Streptomyces filipinensisZQ-22)，分类命名为Streptomyces filipinensis ZQ-22，该菌种已经被中国典型培养物保藏中心-武汉中心保藏，地址：中国.武汉.武汉大学，保藏编号CCTCC NO：M 2015676，保藏日期：2015年11月12日。
本发明提供的放线菌菲律宾链霉菌(Streptomyces filipinensis ZQ-22)是从红树林下土壤中分离的，是具有抗胶质瘤活性的海洋菌株ZQ-22，通过以下步骤分离培养得到：
(1)菲律宾链霉菌(Streptomyces filipinensis ZQ-22)的分离培养
取空气干燥的红树林土壤稀释成一定的浓度，取一定量的样品稀释液均匀分散到含有固体培养基的培养皿中，在室温条件下培养一定时间后,将不同的菌落分别转移到另一含有固体培养基的培养皿中，在室温条件下继续培养一定时间。最后将单一菌落(ZQ-22)接种到斜面培 养基培养后置4℃冰箱保存备用。
所述的红树林土壤是从广东珠海淇澳红树林湿地生态园中获得；所述样品稀释液的浓度为1×10-6～1×10-4g/mL；所述样品稀释液的取样量为100～300μL；所述培养皿所含的固体培养基为高氏琼脂(Gause′s agar)培养基或其它固体培养基；所述的斜面培养基为高氏琼脂培养基或其它固体斜面培养基；所述的室温培养温度为20～30℃；所述的培养时间为7～15天。
(2)菲律宾链霉菌(Streptomyces filipinensis ZQ-22)的菌种鉴定
上述步骤(1)分离培养所获得的菌株ZQ-22使用实验室目前常用的16S rDNA序列分析方法鉴定其种类，确定为菲律宾链霉菌，分类命名为Streptomyces filipinensis ZQ-22。该菌种已经被中国典型培养物保藏中心-武汉中心保藏，保藏编号CCTCC NO：M2015676，保藏日期：2015年11月12日。
本发明的再一个目的是提供一种具有抗胶质瘤活性的化合物Bagremycin C(1)，所述的Bagremycin C为一新化合物，该化合物由Streptomyces filipinensis ZQ-22产生，其化学结构式为：
本发明的第三个目的是提供活性物质Bagremycin C的制备方法，通过以下步骤实现：
(1)菲律宾链霉菌(Streptomyces filipinensis ZQ-22)发酵菌液的制备
取菲律宾链霉菌(Streptomyces filipinensis ZQ-22)的菌落接种到含有一定量的液体菌种培养基的大三角烧瓶中，将含有ZQ-22菌种的培养液在室温条件下振荡培养一定时间后以制备菌种液。最后将菌种液转入含有一定量的液体发酵培养基的大三角烧瓶，在室温条件下振荡发酵培养一定时间后，得到具有抗胶质瘤活性的ZQ-22的发酵菌液。
所述的液体菌种培养基和液体发酵培养基均为液体高氏培养基；所述的用量为100-200mL；所述的大三角培养瓶为250或500mL；所述的室温培养温度为20～30℃；所述振荡的转速为160-180rpm；所述的培养时间为5～15天。
(2)活性物质Bagremycin C的提取分离纯化
上述步骤(1)所获得的菌株ZQ-22发酵菌液过滤后得到菌丝体和发酵液。菌丝体用100％ 甲醇提取，提取液浓缩后得到浸膏A。发酵液通过大孔吸附树脂(DIAION HP-20)柱层析，分别用10％和100％的甲醇洗脱，甲醇洗脱液浓缩后得到浸膏B。浸膏A和浸膏B合并的总浸膏用十八烷基硅烷键合硅胶(ODS)柱层析分离，分别用50％、70％甲醇水和100％甲醇洗脱。70％的甲醇水洗脱液合并后经减压浓缩得到组分C。组分C再用ODS柱层析分离，用60％的甲醇水洗脱，分段收集洗脱液，并用高效液相(HPLC)检测各组分，将含有单一活性物质Bagremycin C的组分合并，减压浓缩后得纯化合物Bagremycin C。
所述柱层析的ODS或DIAION HP-20的用量与上量的样品量比例是40～60g:1.0g；所述的高效液相分离条件是:Agilent 1260高效液相色谱仪，Agilent 1260DAD检测器，Agilent Zorbax SB-C18色谱柱(250×4.6mm,5μm)，75％甲醇/25％水为流动相，柱温26℃,检测波长256nm，流速为1.0mL/min。
(3)活性物质Bagremycin C的结构鉴定
活性物质Bagremycin C的结构是根据它的一维和二维的核磁共振谱(NMR)、高分辨质谱(HRMS)和旋光度[α]D来鉴定的。
本发明的最后一个目的是提供活性物质Bagremycin C在制备治疗胶质瘤药物中的应用。所述的Bagremycin C可显著抑制多种胶质瘤细胞的增殖并诱导肿瘤细胞凋亡；所述药物为Bagremycin C活性成分单独，或Bagremycin C与其它药物或有效成分一起，与药学上可接受的载体组成的药物；所述药物的制剂形式为：液体制剂、固体制剂、胶囊制剂、缓释制剂、纳米制剂。
本发明从红树林下土壤中分离到具有抗胶质瘤活性的海洋放线菌菲律宾链霉菌(Streptomyces filipinensis ZQ-22)，并从该菌株中发现了一种新的抗胶质瘤活性化合物，命名为Bagremycin C。该化合物显著抑制不同胶质瘤细胞的增殖，并显著诱导肿瘤细胞凋亡和影响肿瘤细胞周期。Bagremycin C在制备治疗胶质瘤药物方面具有良好的应用前景。
附图说明
图1.菲律宾链霉菌(Streptomyces filipinensis ZQ-22)的菌落图。
图2-4.活性物质Bagremycin C的氢谱。
图5-7.活性物质Bagremycin C的碳谱。
图8-10.活性物质Bagremycin C的HSQC谱。
图11-13.活性物质Bagremycin C的HMBC谱。
图14.活性物质Bagremycin C的高分辨质谱。
图15.活性物质Bagremycin C的HMBC相关示意图。
图16.活性物质Bagremycin C抑制胶质瘤细胞增殖。
图17.活性物质Bagremycin C阻滞胶质瘤细胞周期于G0/G1期。
图18.活性物质Bagremycin C诱导胶质瘤细胞凋亡。
具体实施方式
以下结合附图和实施例对本发明作进一步详细描述。但是，本发明不限于这些实施例。
实施例1.菲律宾链霉菌(Streptomyces filipinensis ZQ-22)的分离培养
取空气干燥的红树林土壤1克用海水稀释成浓度为1×10-6g/mL的样品稀释液，取200μL的样品稀释液均匀分散到含有高氏琼脂(Gause′s agar)固体培养基的培养皿中，在28℃条件下培养7天后,将不同的菌落分别转移到另一含有高氏琼脂固体培养基的培养皿中，在28℃条件下继续培养7天。最后将生长良好的单一菌落(ZQ-22)接种到高氏琼脂斜面培养基培养后置4℃冰箱保存备用。
实施例2.菲律宾链霉菌(Streptomyces filipinensis ZQ-22)的菌种鉴定
使用16S rDNA序列分析方法鉴定所获得菌株ZQ-22的种类。
2.1实验试剂及仪器
PCR试剂：EX Taq酶(TaKaRa)，dNTP(TaKaRa)，引物(Invitrogen合成)，引物序列是:TACGGYTACCTTGTTACGACTT和AGAGTTTGATCMTGGCTCAG；
Marker：DL5000；
实验仪器：离心机，电泳仪，PCR仪，ABI 3730XL测序仪。
2.2实验步骤
细菌基因组DNA抽提
电泳检测
PCR扩增
a.PCR反应体系
b.PCR反应条件
c.电泳检测
d.测序:切胶纯化测序
e.分析结果:拼接序列。
2.3实验结果
拼接后的序列为：
GGGCCACCGGCTTCGGGTGTTACCGACTTTCGTGACGTGACGGGCGGTGTGTACAAGGCCCGGGAACGTATTCACCGCAGCAATGCTGATCTGCGATTACTAGCGACTCCGACTTCATGGGGTCGAGTTGCAGACCCCAATCCGAACTGAGACCGGCTTTTTGAGATTCGCTCCACCTCACGGTATCGCAGCTCATTGTACCGGCCATTGTAGCACGTGTGCAGCCCAAGACATAAGGGGCATGATGACTTGACGTCGTCCCCACCTTCCTCCGAGTTGACCCCGGCGGTCTCCTGTGAGTCCCCATCACCCCGAAGGGCATGCTGGCAACACAGAACAAGGGTTGCGCTCGTTGCGGGACTTAACCCAACATCTCACGACACGAGCTGACGACAGCCATGCACCACCTGTACACCGACCACAAGGGGGGCACTATCTCTAATGCTTTCCGGTGTATGTCAAGCCTTGGTAAGGTTCTTCGCGTTGCGTCGAATTAAGCCACATGCTCCGCCGCTTGTGCGGGCCCCCGTCAATTCCTTTGAGTTTTAGCCTTGCGGCCGTACTCCCCAGGCGGGGAACTTAATGCGTTAGCTGCGGCACCGACGACGTGGAATGTCGCCAACACCTAGTTCCCACCGTTTACGGCGTGGACTACCAGGGTATCTAATCCTGTTCGCTCCCCACGCTTTCGCTCCTCAGCGTCAGTAATGGCCCAGAGATCCGCCTTCGCCACCGGTGTTCCTCCTGATATCTGCGCATTTCACCGCTACACCAGGAATTCCGATCTCCCCTACCACACTCTAGCTAGCCCGTATCGACTGCAGACTCGGGGTTAAGCCCCGAGCTTTCACAATCGACGTGACAAGCCGCCTACGAGCTCTTTACGCCCAATAATTCCGGACAACGCTTGCGCCCTACGTATTACCGCGGCTGCTGGCACGTAGTTAGCCGGCGCTTCTTCTGCAGGTACCGTCACTCTCGCTTCTTCCCTGCTGAAAGAGGTTTACAACCCGAAGGCCGTCATCCCTCACGCGGCGTCGCTGCATCAGGCTTTCGCCCATTGTGCAATATTCCCCACTGCTGCCTCCCGTAGGAGTCTGGGCCGTGTCTCAGTCCCAGTGTGGCCGGTCGCCCTCTCAGGCCGGCTACCCGTCGTCGCCTTGGTGAGCCATTACCTCACCAACAAGCTGATAGGCCGCGGGCTCATCCTTCACCGCCGGAGCTTTAC AGCCTCCCAGATGCCTGGGAGGCTCATATCCGGTATTAGACCCCGTTTCCAGGGCTTGTCCCAGAGTGAAGGGCAGATTGCCCACGTGTTACTCACCCGTTCGCCACTAATCCACCCCGAAGGGCTTCATCGTCGACTGC。
以上获得的16S rDNA序列与美国NIH的NCBI GenBank数据库比较，其结果表明:菌株ZQ-22的16S rDNA序列与GenBank库中的Streptomyces filipinensis strain NBRC12860的16S rDNA序列有99％的相似性(登录号:KJ789323.1)。因此，本发明所获得的海洋菌株ZQ-22定为菲律宾链霉菌(Streptomyces filipinensis ZQ-22)(附图1)。菲律宾链霉菌(Streptomyces filipinensis ZQ-22)菌株已被中国典型培养物保藏中心-武汉中心保藏，保藏编号CCTCC NO：M 2015676，保藏日期：2015年11月12日。
实施例3.菲律宾链霉菌(Streptomyces filipinensis ZQ-22)发酵菌液的制备
将菲律宾链霉菌(Streptomyces filipinensis ZQ-22)接种到含有200mL液体高氏培养基的500mL三角烧瓶中，将含有ZQ-22菌种的培养液在28℃条件下旋转(180rpm)振摇培养7天后得到菌种液。将5mL菌种液转入含有200mL液体高氏培养基的500mL三角烧瓶中，在28℃条件下旋转(180rpm)振摇培养7天，得到具有抗肿瘤活性的ZQ-22的发酵菌液。
实施例4.活性物质Bagremycin C的提取分离纯化
将上述步骤3所获得的菌株ZQ-22的发酵菌液(25L)过滤后得到菌丝体和发酵液。菌丝体用100％甲醇提取，提取液浓缩后得到浸膏A(1.6克)。发酵液通过大孔吸附树脂(DIAION HP-20，500mL)柱层析，分别用10％和100％的甲醇洗脱，甲醇洗脱液浓缩后得到浸膏B(2.5克)。浸膏A和浸膏B合并的总浸膏用十八烷基硅烷键合硅胶(ODS，300克)柱层析分离，分别用50％、70％甲醇水和100％甲醇各2000mL洗脱。70％的甲醇水洗脱液合并后经减压浓缩得到组分C(1.5克)。组分C再用ODS(100克)柱层析分离，用60％的甲醇水洗脱，分段收集洗脱液，每20mL收集一份，并用HPLC检测各组分，将含有单一活性物质Bagremycin C的组分15-17合并，减压浓缩后得纯化合物Bagremycin C(50.8mg,保留时间为17.7min)。HPLC的检测条件是:Agilent Zorbax SB-C18色谱柱(250×4.6mm,5μm)，75％甲醇/25％水为流动相，柱温26℃,检测波长256nm，流速为1.0mL/min。
实施例5.活性物质Bagremycin C的结构鉴定
Bagremycin C为无色粉末；分子式C20H20N2O6S；[α]D 25-7.67(c 0.50,MeOH)；UV(MeOH)λmax(logε)204(4.97),250(4.84),320(2.78)nm.IR(KBr)3398,2946,1722,1655,1551,1505,1386,1293,1195,1025mm；高分辨质谱(HRESIMS)为m/z[M-H]-415.0970(计算值C20H19N2O6S 415.0964)。
根据Bagremycin C的1H谱(附图2-4)、13C谱(附图5-7)、HSQC谱(附图8-10)、HMBC谱(附图11-13)、高分辨质谱图(附图14)和旋光度，Bagremycin C鉴定为新一化合物，其13C和 1H NMR信号归属见表一,HMBC的相关示意图见附图15。
表一：Bagremycin C的13C谱和1H谱数据
实施例6.Bagremycin C的活性研究
6.1.Bagremycin C抑制胶质瘤细胞增殖的作用
大鼠脑胶质瘤C6细胞和人脑胶质瘤U251细胞用DMEM和10％FBS培养基在37℃和5％二氧化碳的孵化箱中培养，而人脑胶质瘤U87-MG和SHG-44细胞分别用MEM培养基和RPMI-1640培养基在37℃和5％二氧化碳的孵化箱中培养，经过三代培养的细胞用于本发明的实验研究。
用磺酰罗丹明B(SRB)法测定肿瘤细胞存活率，阿霉素(Doxorubicin)用于阳性药对照。细胞接种于96孔板中，贴壁24h后加入不同浓度的Bagremycin C。药物处理72h后用SRB染色，用酶标仪测定515nm处的吸收光度值，检测肿瘤细胞的存活率，计算Bagremycin C抑制胶质瘤细胞增殖的IC50值。实验结果表明：Bagremycin C对四种胶质瘤细胞的增殖均有显著的抑制作用，其IC50值分别为2.21-6.39μM(表二,附图16)。
表二、Bagremycin C对胶质瘤细胞增殖的抑制活性(IC50:μM)
| 化合物 | U251 | U87-MG | SHG-44 | C6 |
| Bagremycin C | 2.21±0.11 | 4.29±0.44 | 2.45±0.23 | 6.39±0.46 |
| 阿霉素 | 3.30±0.73 | 0.42±0.02 | 1.89±0.02 | 0.51±0.07 |
6.2.Bagremycin C阻滞胶质瘤细胞周期于G0/G1期
用碘化丙啶(PI)DNA染色后,流式细胞仪分析Bagremycin C对胶质瘤U87-MG细胞生长周期的影响。将胶质瘤U87-MG细胞分别用Bagremycin C(2.2μM和4.4μM)处理12h和24h后，收集细胞并与冰冻的70％乙醇混合后在4℃条件下过夜。过夜后的混合液经离心(1900rpm，7分钟)分离得到的细胞用PBS洗涤两次。细胞重新分散在含有RNase A的PBS中，在37℃孵化30分钟，最后用碘化丙啶(PI)在4℃黑暗处染色30分钟。用FACScan流式细胞仪分析Bagremycin C对U87-MG细胞周期的改变。
实验结果显示：与对照组比较,Bagremycin C(4.4μM)处理胶质瘤U87-MG细胞12h和24h后,细胞周期G0/G1期的细胞比例分别增加了16.18％和25.21％(表三，附图17)。以上结果说明，Bagremycin C阻滞于胶质瘤U87-MG细胞周期于G0/G1期。
表三、Bagremycin C对胶质瘤U87-MG细胞周期的影响
6.3.Bagremycin C诱导胶质瘤细胞凋亡的作用
用Annexin V-FITC/PI双染色分析方法对Bagremycin C诱导人胶质瘤U87-MG细胞凋亡的作用进行了定量分析。将胶质瘤U87-MG细胞用Bagremycin C(2.2μM和4.4μM)处理24,48和72小时后，收集1×106个细胞。细胞用冷的PBS缓冲液洗涤后重新分散在100μl含有5μl Annexin V-FITC和1μl 100μg/ml PI工作液的结合缓冲液中。细胞在室温下孵化15分钟后加入400μl结合缓冲液，用流式细胞仪检测其荧光(激发波长：488nm；发射波长：530nm和575nm)。实验结果表明：Bagremycin C可显著诱导胶质瘤U87-MG细胞凋亡，其作用与剂量和时间有关。与对照组U87-MG细胞凋亡总数(5.24％)比较，4.4μM的Bagremycin C处理72h后，可引起胶质瘤U87-MG细胞的细胞凋亡总数升高93.0％(表四，附图18，图中左下角为正常细胞，右下角为早期凋亡细胞，右上角为晚期凋亡细胞，左上角为坏死细胞)。
表四、Bagremycin C诱导胶质瘤U87-MG细胞凋亡
以上结果表明：Bagremycin C显著抑制不同胶质瘤细胞的增殖，明显诱导胶质瘤细胞凋亡和影响胶质瘤细胞周期。因此，Bagremycin C在制备抗胶质瘤药物方面具有良好的应用前景。
Claims (5)
1.一种菲律宾链霉菌，其特征在于，其分类命名为Streptomycesfilipinensis ZQ-22，该菌种已经被中国典型培养物保藏中心-武汉中心保藏，保藏编号CCTCC NO：M 2015676，保藏日期：2015年11月12日。
2.一种由权利要求1所述菲律宾链霉菌产生的化合物Bagremycin C，其特征在于，所述的Bagremycin C的化学结构式为：
3.根据权利要求2所述的化合物Bagremycin C的制备方法，其特征在于，通过以下步骤实现：
(1) 菲律宾链霉菌发酵菌液的制备
取权利要求1所述的Streptomycesfilipinensis ZQ-22菌株接种到含有液体菌种培养基的大三角烧瓶中，将含有Streptomycesfilipinensis ZQ-22菌种的培养液在室温条件下振荡培养后制备菌种液，然后将菌种液转入含有液体发酵培养基的大三角烧瓶，在室温条件下振荡发酵培养，得到发酵菌液；
所述的液体菌种培养基和液体发酵培养基均为液体高氏培养基；培养基的用量为100-200 mL；所述的室温培养温度为20-30℃；所述振荡的转速为160-180 rpm；培养所用时间为5-15天；
(2) Bagremycin C的提取分离纯化
上述步骤(1)所获得的发酵菌液过滤后得到菌丝体和发酵液，菌丝体用100%甲醇溶液提取，提取液浓缩后得到浸膏A，发酵液通过DIAION HP-20大孔吸附树脂柱层析，分别用10%和100%的甲醇洗脱，甲醇洗脱液浓缩后得到浸膏B，将浸膏A和浸膏B合并的总浸膏用十八烷基硅烷键合硅胶柱层析分离，分别用50%、70%甲醇水和100%甲醇洗脱，70%的甲醇水洗脱液合并后经减压浓缩得到组分C，组分C再用硅胶柱层析分离，用60%的甲醇水洗脱，分段收集洗脱液，并用高效液相检测各组分，将含有单一活性物质Bagremycin C的组分合并，减压浓缩后得化合物Bagremycin C；
所述柱层析的十八烷基硅烷键合硅胶或DIAION HP-20大孔吸附树脂的用量与上量的样品量比例是40-60 g: 1.0 g；所述的高效液相分离条件是: Agilent 1260高效液相色谱仪，Agilent 1260 DAD 检测器，Agilent Zorbax SB-C18 色谱柱 250
4.6 mm, 5 
，75%甲醇/25%水为流动相，柱温 26℃, 检测波长256 nm，流速为1.0 mL/min；
(3) 活性物质Bagremycin C的结构鉴定
活性物质Bagremycin C的结构是根据一维和二维的核磁共振谱、高分辨质谱和旋光度
D鉴定，确定结构同权利要求2一致。
4.根据权利要求2所述的化合物Bagremycin C在制备治疗胶质瘤药物中的应用。
5.根据权利要求4所述的应用，其特征在于，所述药物为含权利要求2所述的化合物Bagremycin C的活性成分与药学上可接受的载体制成，所述含有Bagremycin C的活性成分选自：1）权利要求2所述的化合物Bagremycin C，或2）权利要求2所述化合物Bagremycin C与其它药物的组成，所述药物的制剂形式为：液体制剂、固体制剂、胶囊制剂、缓释制剂、纳米制剂。
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Non-Patent Citations (4)
| Title |
|---|
| Bagremycin A and B, novel antibiotics from streptomyces sp. Tu 4128;M Bertasso,et al;《J Antibiotics》;20111231;第54卷(第9期);730 * |
| involved in bagremycin biosynthesis in Streptomyces sp Tu 4128.《Annals of Microbiology》.2013,第63卷(第1期),167-172. * |
| Zhu,et al.Cloning and characterization of bagB and bagC, two cotranscribed genes * |
| 新型抗生素Bagremycin生物合成基因的相关研究;朱云霞;《中国优秀硕士学位论文全文数据库》;20140630(第2014/06期);B016-266 * |
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