CN106460021A - 生产葡聚糖的方法 - Google Patents
生产葡聚糖的方法 Download PDFInfo
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- CN106460021A CN106460021A CN201480075195.4A CN201480075195A CN106460021A CN 106460021 A CN106460021 A CN 106460021A CN 201480075195 A CN201480075195 A CN 201480075195A CN 106460021 A CN106460021 A CN 106460021A
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- Prior art keywords
- glucosan
- culture medium
- sucrose
- bacterial strain
- cibarium
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- C—CHEMISTRY; METALLURGY
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- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P19/00—Preparation of compounds containing saccharide radicals
- C12P19/04—Polysaccharides, i.e. compounds containing more than five saccharide radicals attached to each other by glycosidic bonds
- C12P19/08—Dextran
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- A—HUMAN NECESSITIES
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- A23C19/00—Cheese; Cheese preparations; Making thereof
- A23C19/02—Making cheese curd
- A23C19/05—Treating milk before coagulation; Separating whey from curd
- A23C19/054—Treating milk before coagulation; Separating whey from curd using additives other than acidifying agents, NaCl, CaCl2, dairy products, proteins, fats, enzymes or microorganisms
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L29/00—Foods or foodstuffs containing additives; Preparation or treatment thereof
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- A23L29/273—Dextran; Polysaccharides produced by leuconostoc
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- C—CHEMISTRY; METALLURGY
- C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
- C08B—POLYSACCHARIDES; DERIVATIVES THEREOF
- C08B37/00—Preparation of polysaccharides not provided for in groups C08B1/00 - C08B35/00; Derivatives thereof
- C08B37/0006—Homoglycans, i.e. polysaccharides having a main chain consisting of one single sugar, e.g. colominic acid
- C08B37/0009—Homoglycans, i.e. polysaccharides having a main chain consisting of one single sugar, e.g. colominic acid alpha-D-Glucans, e.g. polydextrose, alternan, glycogen; (alpha-1,4)(alpha-1,6)-D-Glucans; (alpha-1,3)(alpha-1,4)-D-Glucans, e.g. isolichenan or nigeran; (alpha-1,4)-D-Glucans; (alpha-1,3)-D-Glucans, e.g. pseudonigeran; Derivatives thereof
- C08B37/0021—Dextran, i.e. (alpha-1,4)-D-glucan; Derivatives thereof, e.g. Sephadex, i.e. crosslinked dextran
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- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
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- C12N1/20—Bacteria; Culture media therefor
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- C12Y—ENZYMES
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- A—HUMAN NECESSITIES
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- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
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- Chemical Kinetics & Catalysis (AREA)
- Molecular Biology (AREA)
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- Dispersion Chemistry (AREA)
- Nutrition Science (AREA)
- General Chemical & Material Sciences (AREA)
- Materials Engineering (AREA)
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Abstract
一种生产葡聚糖的方法,包括如下步骤:制备包含合适的混合物和成分平衡的培养基,主要经过对碳和氮源的性质和浓度的准确选择,且采用特定初始pH,对培养基用合适的细菌株(将生产标准化并尽可能避免系统的变化)的量进行接种;以规定的时间和规定的温度进行发酵;将葡聚糖沉淀以从培养基中分离产物;所述细菌株是食窦威克斯氏菌菌株。
Description
技术领域
本发明涉及一种用于生产葡聚糖的方法,具体地,涉及一种葡聚糖的优化的生物合成方法。
背景技术
葡聚糖是一种由葡萄糖单元形成的多糖,其链延长通过葡聚糖蔗糖酶来催化。葡聚糖是一种可变侧链1-3与葡聚糖聚合物的主链单元连接的α-D-1,6-葡糖连接的葡聚糖;该产品具有对最终溶液有影响的不同的分子量(≥1000Da)。天然葡聚糖粉末的化学和物理特性根据生产它的生产微生物株和/或生产方法而改变。葡聚糖的生物合成已在许多细菌中被证实,尤其在变形链球菌(Streptococcus mutans)、肠膜明串珠菌肠膜亚种(Leuconostoc mesenteroides ssp.mesenteroides)和肠膜明串珠菌葡聚糖亚种(Leuconostoc mesenteroides ssp.dextranicum)。明串珠菌属在蔗糖的存在下生产葡聚糖蔗糖酶并将其分泌至培养基内。该酶,即葡聚糖蔗糖酶,自蔗糖基质合成葡聚糖,催化葡糖基残基从蔗糖向葡聚糖聚合物的转移并释放果糖。葡聚糖蔗糖酶的来源(即其生产微生物)影响着葡聚糖分子的分支点的出现次数和性质。
葡聚糖是一种易溶的、生物相容且生物可降解的聚合物;市售的天然葡聚糖粉末已被应用于多个领域。它在生物化学中尤其被作为Sephadex型的凝胶上的过滤色谱的载体使用。葡聚糖可用于化妆品工业和药物组合物中(例如参见专利US5902800)。而且,在治疗领域中,它被用作血浆的替代品(《普通生物化学》(Biochimie generale(GeneralBiochemistry))-J.H.WEIL-Masson,第六版-1990-第171页)。另外,通过葡聚糖明串珠菌的菌株合成的葡聚糖被用于食品工业,以获得食品如酸奶、奶油甜品、奶基饮品和色拉酱的质地。欧洲专利申请公开第EP0363633号证实了通过葡聚糖明串珠菌的菌株,具体是葡聚糖明串珠菌的NRRL-B-18242菌株的葡聚糖的合成。此外,该专利申请的公开特别记载了一种包含通过该细菌合成的葡聚糖的组合物和该组合物在食品领域中的应用。葡聚糖的食品应用遵循着顾客希望真实、美味和天然地制造食物并避免化学添加剂的趋势。槽式发酵而得的天然添加剂满足了食品生产者对选择天然成分的要求,其实现安全、可靠和可持续。葡聚糖粉末还可作为织构剂用于烘培,主要在无麸质面包中,增强最终产品的技术性能。以此为目的,高分子量葡聚糖(1-2·106Da)已被欧盟批准用作烘培产品中的食品成分(Naessens M.等,2005)。
最近,对可实现高分子量葡聚糖的高产率的细菌的寻找已找到一种称为食窦威克斯氏菌(Weissella cibaria)的菌种。食窦威克斯氏菌是一种革兰氏阳性的异型发酵菌菌种,属于明串珠菌科,在2002年《国际系统和进化微生物杂志(International Journal ofSystematic and Evolutionary Microbiology)》52(Pt 1):141–8)的《融合威克斯氏菌的分类研究以及食品和临床样本中发现的食窦威克斯氏菌新种的描述(Taxonomic study ofWeissella confusa and description of Weissella cibaria sp.nov.,detected infood and clinical samples)》中被定义(KJ、Schillinger U、Geisen R等,2002年1月)。食窦威克斯氏菌被美国食品药品管理局(FDA)认为是GRAS细菌(通常认为安全),且该属也被包括在欧洲食品安全局提议的分类学单元列表中(EFSA,QPS表,安全资格认证(Qualified Presumption of Safety))。鉴于许多工业应用,该株非常重要。该菌种从天然基质中被分离,并在由蔗糖形成粘性物质之后被选择。还发现其在由蔗糖进行的葡聚糖合成中是高产的。
发明目的
得到本发明的研究的第一个目的是从天然食品基质分离微生物并鉴定能够以高产率生产葡聚糖的这一细菌菌株,尤其是来自食窦威克斯氏菌的菌株。本发明的另一个目的是提供一种能够以高产率生产高分子量葡聚糖粉末的生产葡聚糖的定制方法。
因此,本发明的一个目的是一种生产葡聚糖的定制方法,包括以下步骤:
-制备优化的合成培养基,该培养基包含正确平衡的营养物质,特别是选择(考虑性质和浓度方面)合适的碳和氮源,(在该方法的微调试验之后)具有给定的pH;
-采用合适的(考虑寿命和用量方面)的细菌预培养物的接种物大小来确保良好的生长速率(在达到指数期后冻干以使过程标准化);
-在给定的最佳温度下以给定的时间进行孵育(因为高温会减弱细胞生长并导致酶的局部不稳定)。
-从优化了产品的下游回收和增加到最大产率的培养基中分离合成的葡聚糖。
以上所有描述的步骤是针对保藏号NCIMB 42196(2013年11月)的食窦威克斯氏菌细菌株进行优化的。该菌株是非产芽孢细菌、非运动性、微好氧菌、异型发酵菌和过氧化氢酶阴性,由L-阿拉伯糖而非半乳糖产生酸。
本发明的一个目的是用于生产高分子量葡聚糖的基于保藏号NCIMB 42196的食窦威克斯氏菌株。
在本发明的优选的实施方式中,用于葡聚糖生产的最优氮源是酵母提取物,以百分比为约1%到2%w/v。碳源主要是蔗糖,以百分比为10%到15%w/v。
在本发明的另一个实施方式中,培养基还包括富集的斯科特肉汤(scotta-broth),或奶酪工业的类似副产品,以百分比为约80%到90%。斯科特肉汤是一种可变基质,基本由盐和矿物质(在乳清奶酪制造工艺后残留)制成。该天然食品基质的组合物根据生产步骤和原材料(牛乳)特性而改变。
培养基的初始pH值较好地调整为pH6-7左右,优选约pH6.5。
孵育时间为20到36小时,优选24小时。孵育在约50rpm的轻柔搅拌下进行。
孵育在约28℃到32℃下进行,优选30℃。
另一个目的是通过上述食窦威克斯氏菌株的细菌而生产的葡聚糖蔗糖酶。所述葡聚糖蔗糖酶的基因组序列和蛋白质序列已被检测和列出,并被附于本申请。
本发明的另目的在于一种根据如上所述的方法而得的高分子量(5·106到4·107Da之间)葡聚糖粉末。根据本发明的方法而得的葡聚糖粉末具有7%到11%的蛋白质含量,且主要为9%,而且葡聚糖溶液的特征粘度值为4.0到5.0mPa·s(在20℃-25℃的温度下)。
发明实施方式描述
文献示出了影响微生物的生物合成的各种工艺参数导致了葡聚糖生产变化性的许多例子。分离具有潜在工业应用前景的生产葡聚糖的微生物和鉴定影响葡聚糖生产的因素的最佳组合是本工作的两个主要的焦点。
为了使用合适的培养基组合物(考虑必需的营养需求和合适的变量方面)和优化的工艺参数(考虑使用特定乳酸细菌株的工业规模生产方面)来得到高产率,在振荡烧瓶(500ml)和批式发酵(不进行pH控制)的条件下进行实验。
在所有的实验中,使用在MRS培养基中于30℃下生长18-20小时后的我们的根据保藏号NCIMB 42196(6·107CFU/ml)的食窦威克斯氏菌的冻干菌株接种物(加入量:1/200w/v),并通过乙醇沉淀并在100℃下干燥来测定葡聚糖。
实施例1.氮源和浓度对葡聚糖生产的影响。
保持恒定的蔗糖浓度(10%w/v),其目的是验证葡聚糖生产是否受氮(和其他的盐)供应的影响。在测试了富含磷酸盐和氮源的一些培养基、这些营养物质类型(或它们的组合)的浓度和其他欠佳方面之后,我们发现葡聚糖生产敏感地受氮源影响,且酵母提取物是最佳的营养源(在经测试的情况中)。考虑到酵母提取物由酵母细胞(酿酒酵母,Saccharomyces)的自溶而得,并且它是氨基-氮和维生素,尤其是水溶性复合维生素B的优良来源,其确保了在极短时间内的良好细胞生长(不论其他经测试的来源)。另外,将酵母提取物和其他盐组合(参考其他示例),可给予最佳的营养平衡以促进细胞增殖。
培养基1a(蛋白胨1%w/v,蔗糖10%w/v)
培养基1b(蛋白胨2%,蔗糖10%)
培养基2a(酵母提取物1%,蔗糖10%)
培养基2b(酵母提取物1.5%,蔗糖10%)
培养基2c(酵母提取物2%,蔗糖10%)
培养基3(硝酸铵1%,蔗糖10%)
培养基4(硫酸铵1%,蔗糖10%)
培养基5(氯化铵0.5%,硝酸钾0.5%和硝酸钠0.5%,蔗糖10%)
不同的氮源(简单盐或复杂基质)不允许相同的葡聚糖生产(以最终产率计),且葡聚糖的最高量与酵母提取物(1%到2%,最大蔗糖转化百分比在1.5%下产生)的引入有关,其也增加了细胞生长(减少生产时间)。换而言之,约1.5%的酵母提取物浓度揭示了细菌细胞生长和形成产物之间的最佳平衡(在其他实验中,该基础培养基用其他营养源富集来使产率最大化)。
实施例2.碳源的性质和浓度对葡聚糖生产的影响。
保持所选氮源(酵母提取物)固定,其目的是验证不同碳源(代替蔗糖)对葡聚糖生产的影响。在各培养基中加入5%w/v的蔗糖。蔗糖与如下替代碳源一起加入:玉米浆、葡萄糖、果糖、甘露糖、乳糖(在各培养基中加入1.5%w/v的酵母提取物)。
培养基1:玉米浆5%(1a)和10%(1b)+蔗糖5%
培养基2:葡萄糖10%w/v+蔗糖5%w/v
培养基3:甘露糖10%+蔗糖5%
培养基4:乳糖10%+蔗糖5%
在存在不同碳源以代替蔗糖的情况下,葡聚糖生产总是且无差别地较低。该菌株使用蔗糖作为用于葡聚糖生产的唯一碳水化合物来源(如关于其它菌种例如肠膜明串珠菌(L.mesenteroides)所报道的——Cavenaghi,2000)。相对于其它经测试的碳源,蔗糖似乎是葡聚糖生产的诱导物(归因于特定酶的诱导)。而且将两种不同的碳源混合也不会明显增加葡聚糖的生产。
实施例3.蔗糖浓度对葡聚糖生产的影响。
保持酵母提取物浓度(1.5%w/v)恒定并使用蔗糖作为唯一可获得的碳源,其目的是确定基质浓度对葡聚糖生产的影响。
培养基1:5%w/v蔗糖
培养基2:10%蔗糖
培养基3:15%蔗糖
培养基4:20%蔗糖
培养基5:25%蔗糖
*高残留蔗糖
初始蔗糖浓度越高,得到的每单位体积葡聚糖的产率越高。结果是,生长率、葡聚糖生产和转化时间(还考虑蔗糖的转化百分比,没有基质残留)之间的最佳平衡在使用10-15%(w/v)蔗糖时得到。最佳实验条件(pH6.5,温度30℃,酵母提取物1.5%w/v并添加其他盐,合适的接种物大小)下的最大特定生长率(μ最大)预计是约0.94h-1。
实施例4.初始pH对葡聚糖生产的影响。
将MRS培养基(补充蔗糖直至最终浓度为15%w/v)用于这些实验。考虑对细胞生长和最终的葡聚糖生产的影响,最佳的初始pH(灭菌前,用NaOH1M调整)为6.0–7.0(最佳结果在6.5获得)。
培养物的最终pH(在发酵终点时)是约3.5。
实施例5.搅拌速度(搅动)对葡聚糖生产的影响。
将含有MRS培养基(补充蔗糖直至最终浓度为15%w/v)的烧瓶用于这些实验。一些实验在使用不同的搅拌速度(50、100、150、200、250、300rpm)的情况下进行。结果发现,搅拌速度没有对葡聚糖生产产生较大影响,因此为了降低起泡风险以及在工艺过程中节省能源,将最佳搅拌速度选为50rpm。
该菌株是兼性微好氧菌,且实验证据证实氧供给对菌株生长有积极影响但对生产葡聚糖没有显著影响。发酵实验中所使用的有氧条件(在20L的生物反应器中)是约1.0mmol/l·h的氧转移速率。
实施例6.接种物大小对葡聚糖生产的影响
将有MRS培养基(补充蔗糖直至最终浓度为15%w/v)的烧瓶用于这些实验。
在所有实验中使用在合成培养基(蔗糖10-15%w/v,酵母提取物1.5%w/v,K2HPO40.4%w/v,乙酸钠3H2O 1%w/v,柠檬酸0.4%w/v,MgSO4·7H2O 0.05%w/v)中在30℃下生长18-20小时后的我们的冻干菌株的接种物(6·107CFU/ml)(加入的接种物的量:1/100w/v、1/200w/v、1/250w/v、1/300w/v)。
接种物大小 | 葡聚糖(g/L) |
1 | 49.5±0.1 |
2 | 58.3±0.07 |
3 | 60.2±0.2 |
4 | 48.8±0.1 |
接种物大小主要影响发酵时间,并且最佳实验结果(考虑细胞生长的标准化、发酵时间和葡聚糖生产)在使用稀释度为1/200w/v的食窦威克斯氏菌菌株的冻干细胞时获得。
实施例7.孵育时间对葡聚糖生产的影响。
将有MRS培养基(补充蔗糖直至最终浓度为15%w/v)的烧瓶用于这些实验。为了确定葡聚糖生产,需考虑细菌细胞已度过迟缓期、适应于培养基且在可获得碳源(和其他营养)的情况下一直生长。由于这些原因,在16到36小时内注意孵育时间以寻得最佳的葡聚糖生产。
发现24小时(至多36小时)的孵育时间是最佳的孵育时间。总之,应通过如下双重检查来控制生产工程:发酵中培养基的粘度的上升和pH的下降。
最终复合和合成的培养基组合物(水中),以使生长速率最大化并使葡聚糖生产保持最高标准(最少24/36小时中):
温度:30℃
发酵时间:24小时(最大36小时)
接种物大小:1/200w/v冻干细胞(6·107CFU/ml,在MRS培养基中在30℃下18-20小时后)
葡聚糖是中性且可溶于水的多糖,因此pH或盐浓度的改变不显著影响粘度。葡聚糖是一种大尺寸的中性聚合物,因此它不易通过/扩散通过人体细胞和组织,可保持良好的渗透压。进行(流变仪的的底板上的)动态流变实验,并检测葡聚糖-水溶液(不同浓度下,pH6.5)的粘度(由于理想粘性液体的特性,所有溶液的粘度在剪切速率上是独立的),15%葡聚糖-水溶液的最终粘度为约210η(mPa*s)且1%葡聚糖最终溶液的最终粘度为约5η(mPa*s)。
另一个高分子量葡聚糖的可能的食品应用是参与奶酪生产,并且这基于葡聚糖是良好的脂肪替代品的特性。已知许多市售脂肪替代品(例如基于乳清蛋白、淀粉和黄原胶或微晶纤维素)具有制造更优的低脂肪产品的潜力;其中的大多数基于微粒化材料且要求高生产成本。
将相同的食窦威克斯氏菌(保藏号NCIMB 42196)菌株用于在基于斯科特肉汤、富含蔗糖和其他的盐的合成培养基中接种。该计划的第二部分的目的是回收乳品业的副产品,以避免去除副产品的成本并提高食品产品质量。斯科特肉汤是一种从乳清乳酪生产工艺中分离的基质,其是盐和营养组成方面可变的副产物。
斯科特肉汤通常包括低水平的蛋白质(0.10-0.15%)、高浓度的盐(0.9-1.2%)和有机酸(0.20-0.25%);脂肪约0.15-0.30%以及低水平的残留乳糖。基于斯科特肉汤(如下所示,富含蔗糖和酵母提取物)的发酵合成培养基,能够获得一种粘性天然发酵液体,该液体又可进一步被包括在奶酪制作生产中,并被称为葡聚糖-糊剂(在发酵中自然地在葡聚糖中以最终浓度8-10%富集)。这可成为提高该副产物的价值和富集最终产品中健康特性的契机(不改变实际工艺中的任何一个步骤)。
在奶酪生产(具有粘度约600-700cp的特征,归因于在发酵中自然累积葡聚糖)中简单地将经发酵的葡聚糖-糊剂加入到原料乳中,能够增加生产的产率并获得低脂肪奶酪(直至最终脂肪浓度4-5%,按照美国食品标签要求中报告低脂肪食品的参照量为≤3g脂肪/50g)。
根据清洁标签(clean labeling)的概念(没有任何关于其他食品添加剂的添加的声明)应将经发酵的葡聚糖-糊剂直接并入奶酪基质,并且其与影响奶酪结构分布的酪蛋白相互作用。由于葡聚糖制造的水的结合可提高奶酪的水含量,因而可改善低脂肪奶酪的性质和性能。奶酪的脂肪含量影响产品的微观结构和高含水量。
用于葡聚糖糊剂的培养基组合物:
蔗糖 10-15%w/v
酵母提取物 1-1.5%w/v
斯科特肉汤 83-89%w/v
关于参与葡聚糖合成的酶:
葡聚糖蔗糖酶或GH 70是一种糖苷水解酶家族70的胞外酶,其对葡萄糖之间的糖苷键进行切割并常常结合碳水化合物模块。该酶以单个或多个分子的形式存在,且具有不同的分子量。金属离子如Ca2+、Mg2+和Co2+可提高酶活性,其他金属离子如Cu2+、Fe3+和Mn2+抑制葡聚糖蔗糖酶活性(Kobayashi M.和Matsuda K.,1976:Goyal A.,Nigam M.和KatiyarS.S.,1995)。
葡聚糖蔗糖酶的基因组序列已通过根据保藏号NCIMB 42196的食窦威克斯氏菌菌株被检测和列出,并被附于本申请。
PCT/RO/134表
Claims (14)
1.一种用于生产葡聚糖的方法,包括以下步骤:
a)制备包含合适的混合物成分平衡的培养基,主要经过对碳和氮源的性质和浓度进行准确选择,并且采用特定初始pH;
b)用合适量的细菌株(以使生产标准化并尽可能避免系统的变化)接种培养基;
c)以给定的时间和给定的温度进行发酵。
d)使葡聚糖沉淀以从培养基中分离产物;
其特征在于,所述细菌株是来自保藏号NCIMB 42196的食窦威克斯氏菌(Weissellacibaria)菌株。
2.如权利要求1所述的方法,其特征在于,所述氮源是酵母提取物,其百分比为约1%到2%w/v;所述碳源包括蔗糖,其百分比为约10%到15%w/v。
3.如权利要求1或2所述的方法,其特征在于,所述培养基还包括斯科特肉汤,或奶酪工业的类似副产品,其百分比(w/v)为约80%到约90%。
4.如权利要求3所述的方法,其特征在于,所述斯科特肉汤包括低水平的蛋白质(0.10-0.15%)、高浓度的盐(0.9-1.2%)和有机酸(0.20-0.25%);脂肪为约0.15-0.30%,和低水平的(4.0-4.6%)残留乳糖。
5.如权利要求1~4中任一项所述的方法,其特征在于,所述培养基的初始pH值较好地被调整为pH6-7附近,且优选为约pH6.5。
6.如权利要求1~5中任一项所述的方法,其特征在于,孵育时间为20到36小时之间,优选24小时。
7.如权利要求1~6中任一项所述的方法,其特征在于,孵育在约50rpm的轻柔搅拌下进行。
8.如权利要求1~7中任一项所述的方法,其特征在于,孵育优选在约25℃到35℃下进行,且最优温度为30℃。
9.一种根据保藏号NCIMB 42196的用于生产葡聚糖的食窦威克斯氏菌菌株。
10.通过如权利要求9所述的食窦威克斯氏菌菌株生产的葡聚糖蔗糖酶。
11.根据权利要求1~8中任一项所述的方法生产的葡聚糖,所述葡聚糖具有5·106到4·107Da的平均分子量,且蛋白质含量为7%到11%,通常为9%。
12.如权利要求11所述的葡聚糖,其特征在于,其粘度为4.0到5.0mPa·s(在20℃-25℃的温度下)。
13.一种葡聚糖糊剂,其特征在于,其包含至少8-10%的葡聚糖,所述葡聚糖根据权利要求3~8所述的方法生产。
14.一种低脂肪奶酪,其特征在于,所述低脂肪奶酪通过在同一发酵工艺过程中将权利要求13所述的葡聚糖糊剂加入原料乳来生产。
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