CN107858310A - 一株产香放线菌及其对诺邓火腿增香的应用 - Google Patents
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Abstract
本发明属于微生物技术领域，涉及一株产香放线菌及其对诺邓火腿增香的应用。所述菌株为耐盐高温双歧菌（Thermobifida halotolerans），分离自诺盐盐矿，菌株保藏号为YIM Yu1701。本发明将耐盐高温双歧菌作为发酵剂，在诺邓火腿的腌制阶段进行接种，火腿成品挥发性成分的总类数显著增加，增香效果明显。
Description
技术领域
本发明属于微生物技术领域，具体涉及一株产香放线菌及其对诺邓火腿增香的应用。
背景技术
诺邓火腿是云南省云龙县诺邓村的特产，与宣威火腿、鹤庆的圆腿并称为云南三大名腿。诺邓火腿传承着用盐泥敷腌火腿的千年工艺，生产出的火腿香味浓郁、色泽红润、入口美不胜收、满口余香，相传南诏时期即是宫廷贡品，千百年来已成为远近闻名的地方名优土特产品。
腌制诺邓火腿的盐，是本地自产自销的诺盐，再加上诺邓特殊的气候条件，造就了诺邓火腿优良的品质。本发明提供的产香放线菌分离自诺盐盐矿。经鉴定，该放线菌为耐盐高温双歧菌（Thermobifida halotolerans），具有独特的产香功能。该耐盐高温双歧菌是云南大学微生物研究所2008年发现的高温双歧菌属的一个新种，目前对它的研究主要集中在酶学方面（产高温纤维素酶和木聚糖酶等），对其挥发性产物的研究未见报道。本发明将耐盐高温双歧菌作为发酵剂，在诺邓火腿的腌制阶段进行接种，火腿成品挥发性成分的总类数显著增加，增香效果明显。
发明内容
本发明的目的之一在于提供一株产香放线菌菌株。
本发明的另一目的在于提供上述菌株在诺邓火腿腌制阶段，使火腿增香的应用。
菌株及其特性：
本发明提供的产香放线菌菌株为耐盐高温双歧菌（Thermobifida halotolerans），已于2017年04月18日保藏于云南省微生物研究所，菌株保藏号为YIM Yu1701。该菌株的特性如下。
好氧，革兰氏阳性，在ISP培养基2、PDA和营养琼脂培养基中生长良好，在ISP培养基3、4和5中生长缓慢，在查氏培养基中不生长。基内菌丝体发达，气生菌丝体仅在营养琼脂培养基上形成。孢子光滑、单个着生于气生菌丝上，气生菌丝呈二叉分枝状不断延伸。其最适生长条件为：温度45℃，pH7.0-8.0。硝酸盐还原试验呈阳性，能产生纤维素酶、淀粉酶以及木聚糖酶等，明胶液化阴性，牛奶胨化和凝固阴性，H2S试验和脲酶试验阴性，不产生黑色素。
对上述菌株进行16S rDNA序列分析，结果显示，它与耐盐高温双歧菌模式菌株YIM90462T相似度达到100%。
菌株对诺邓火腿增香应用：
先将上述菌株进行安全性检测，包括溶血性试验和动物试验，确定其为无毒型菌株。然后将其作为发酵剂，在诺邓火腿的腌制阶段进行接种，待火腿成熟后利用固相微萃取技术提取挥发性成分，利用气相色谱-质谱( GC-MS) 联用技术分析其组成。
附图说明
图1表示的是菌株对于诺邓火腿的发酵工艺。
具体实施方式
以下实施例可以对本发明作进一步说明：
1.菌株的安全性检测
1.1 溶血性试验：以普通营养琼脂培养基为基础，加入羊血混匀，趁热倾注平皿，冷却保藏，将被检菌划线接种于血琼脂平板中，经37℃倒置培养48h后，观察其菌落周围是否产生溶血圈。
1.2 动物试验：将所要检验的菌株活化两代，用生理盐水调节菌体浓度为106cfu/mL。用无菌注射器吸取菌液1mL注射于小鼠腹腔，观察小鼠饮欲、食欲、活动及死亡情况，试验期为两周。
2.菌株的安全性检测结果
经溶血性试验，菌落周围未产生溶血圈。在动物试验期间，小鼠生长良好，进食正常，未见死亡。由此可推断其为无毒型菌株。
接种：将上述菌株作为发酵剂，在诺邓火腿的腌制阶段进行接种。
3.香气成分提取及GC-MS分析
以不接种菌的诺邓火腿为对照组，利用固相微萃取技术对火腿成品进行香气成分的提取，利用气相色谱-质谱（GC-MS）联用技术分析其组成。
4.1 香气成分提取：将固相微萃取头在气相色谱进样口250℃老化2h。火腿样品冷冻24h后切成细粒，取4g放入15 mL样品瓶中，萃取头插入样品瓶并推出吸附纤维头，60℃恒温吸附40min，随后插入气相色谱仪进样口250℃解吸2min。
4.2 香气物质的鉴定：采用HP6890型气相色谱/5973型质谱联用仪对香味成分进行分析和鉴定。SPME纤维头上所吸附的挥发性化合物被解吸下来，挥发性化合物在HP-5MS色谱柱上完成分离（以N2为载气，流速是1.0 mL/min）。
5. 香气成分（挥发性成分）统计分析
用质谱检索化合物时，由于化合物总数太多，所以只对相对百分含量≥0.01%的化合物进行检索。风味是火腿最重要的质量指标，而在火腿腌制过程中，脂肪、蛋白质等风味前体物质在各种因素的作用下发生了酶促或非酶变化，形成了醛类、酸类、酯类等挥发性成分，是火腿风味香气的重要来源。由表1可知，相对于不接菌的对照组，接种耐盐高温双歧菌的诺邓火腿其挥发性成分的总类数显著增加。这表明耐盐高温双歧菌对诺邓火腿的增香效果明显。
表1 不同处理的诺邓火腿挥发性成分的种类比较
| 处理 | 醛类 | 酸类 | 醇类 | 烃类 | 酯类 | 酮类 | 其他 | 总计 |
| 接菌 | 13 | 11 | 4 | 5 | 7 | 5 | 19 | 64 |
| 不接菌 | 12 | 8 | 6 | 5 | 5 | 3 | 10 | 49 |
Claims (3)
1.一株产香放线菌菌株，其特征在于，所述菌株为耐盐高温双歧菌（Thermobifida halotolerans），菌株保藏于云南省微生物研究所，菌株保藏号为YIM Yu1701。
2.根据权利要求1所述的耐盐高温双歧菌菌株，其特征在于：耐盐高温双歧菌菌株好氧，革兰氏阳性，在ISP培养基2、PDA和营养琼脂培养基中生长良好，在ISP培养基3、4和5中生长缓慢，在查氏培养基中不生长；基内菌丝体发达，气生菌丝体仅在营养琼脂培养基上形成；孢子光滑、单个着生于气生菌丝上，气生菌丝呈二叉分枝状不断延伸；其最适生长条件为：温度45℃，pH7.0-8.0；硝酸盐还原试验呈阳性，能产生纤维素酶、淀粉酶以及木聚糖酶等，明胶液化阴性，牛奶胨化和凝固阴性，H2S试验和脲酶试验阴性，不产生黑色素。
3.根据权利要求1所述耐盐高温双歧菌菌株的应用，其特征在于：将耐盐高温双歧菌菌株作为发酵剂在诺邓火腿的腌制阶段进行接种，火腿成品挥发性成分的总类数显著增加，增香效果明显。
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|---|---|---|---|---|
| CN114591863A (zh) * | 2022-03-22 | 2022-06-07 | 云南大学 | 用于提高断奶仔猪生长性能和免疫功能的藏猪源菌剂组合 |
| CN114591863B (zh) * | 2022-03-22 | 2022-12-20 | 云南大学 | 用于提高断奶仔猪生长性能和免疫功能的藏猪源菌剂组合 |
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