CN86104370A - 杂多糖s-184 - Google Patents
杂多糖s-184 Download PDFInfo
- Publication number
- CN86104370A CN86104370A CN86104370.7A CN86104370A CN86104370A CN 86104370 A CN86104370 A CN 86104370A CN 86104370 A CN86104370 A CN 86104370A CN 86104370 A CN86104370 A CN 86104370A
- Authority
- CN
- China
- Prior art keywords
- alcaligenes
- mixed polysaccharide
- carbohydrate
- fermentation
- substratum
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Granted
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Abstract
新杂多糖S-184是用新的产碱杆菌菌种ATCC53160通过发酵制取。它在水溶液中具有可作为增稠剂、悬浮剂和稳定剂的重要性质,用于食品中特别有价值。
杂多糖S-184主要是由碳水化合物组成,还含有12%到16%的蛋白质和3.0%到4.5%酰基(以醋酸计算),碳水化合物部分包含约7%到16%的糖醛酸和中性糖:甘露糖,葡萄糖和半乳糖,它们的克分子比大致为1∶3∶5。
Description
这发明属于微生物多糖范围。在这领域中已知某些微生物的共同特点是产生胞外杂多糖,杂多糖具高分子量,通常由二种或多种单糖组成重复单元聚合而成直链碳水化合物的聚合物。
极大数杂多糖的有用性是基于它们有改变水溶液的粘度和流变学的能力。此外杂多糖还有次级功能和乳化作用,悬浮作用,稳定作用,絮凝作用,润滑作用,成膜作用等。
杂多糖是广泛用于食品,钻机,农业和其他多种工业。近几十年来对这些水溶性胶在商业上的需求量大大增加了。此外各种新的工业技术产生了对具有新物理性质的杂多糖的需求。因此,对杂多糖不同功能范围的需要结合到商业上的需求,已明显地说明发展新的和具有不同物理性质的新杂多糖的必要性。
所以本发明的目的是提供一个由产碱杆菌属新种产生的新杂多糖。此外本发明又可提供制作这个新杂多糖的方法。另一个目的是提供为生产这个新化合物的微生物。更进一步的目的是探索含有此新杂糖的组成。本发明的这些以及其他目的,从以下的描述中将会很清楚的。
现已找到一种新的杂多糖,它的主要组分是碳水化合物,还含有12%-16%的蛋白质,3.0%-4.5%的酰基(以醋酸计算),碳水化合物部分大致含7%到16%的糖醛酸和各种中性糖:甘露糖、葡萄糖和半乳糖的克分子比大致是1∶3∶5;是由产碱杆菌的一个新种,在选择过的碳源上产生的。这新型化合物是用一个未定种名的产碱杆菌在合适的营养培养基中,通气发酵产生的,按照布达佩斯条约,这菌的生物学纯菌株由马里兰州洛克菲尔市,美国典型菌株收集中心保存,1985年6月19日进入登记号ATCC53160菌株。这个杂多糖是以杂多糖S-184表示,在水的体系中有理想的性质,尤其是用于食品中。
本发明的新型菌株是从加利福尼亚州圣地亚哥帕罗玛山的波玛。格林克收集的水样中分离到的。该菌从酵母肉汁琼脂平板,30℃培养4天后的胶粘菌落挑出。然后在培养琼脂上进行纯培养。
种子瓶由分离株的营养琼脂培养物开始。这个种子以后用于接种另一个含有以水解淀粉作炭源的营养培养基的瓶。培养以后,这瓶表现出一定的粘稠,加入异丙醇就沉淀出纤维状物质。开始进行另一瓶种子,并用以测定各种营养培养基对产胶的影响和确定此菌。最佳生长培养基以及发酵条件。
杂多糖S-184是经ATCC53160菌接种在合适的营养培养基中在控制条件下通气发酵产生的。这培养基含有可同化的炭源、氮源和无机盐。
通常在营养培养基中碳水化合物(如葡萄糖、果糖、麦芽糖、木糖及类似物)能单独亦能组合作为可利用的炭源。在培养基中碳水化合物作单一源或组合源的利用精确量,部分是根据培养基中其他组分而定,但一般碳水化合物的量大约为培养基重量的2%和5%之间变动。这些碳源在培养基中可以单独使用亦可组合使用。
通常很多蛋白质类物质可用作发酵过程中的氮源。合适的氮源包括如酵母水解物,初级酵母,黄豆饼粉,棉籽饼粉,水解酪素,玉米浆,蒸馏可溶物或蕃茄酱等等。氮源既可单独亦可组合应用,其用量大约为培养重量的0.05%到0.2%范围。
在培养基中能掺合的营养无机盐,是常用的盐类,能产生钠,钾,铵,钙,磷酸根,硫酸根,氯化物,碳酸根等离子。还包括微量金属如钴,镁,铁和锰。
必须指出,在这里所描述的营养培养基仅仅是说明可能应用的多种多样培养基,但并不限于这些。
作为一个另一可替代的培养基,S-184菌可以在低浓度钙离子条件下生长,例如在无离子水或其他基本不含钙离子的水溶液系统(例如在最终发酵液中少于4ppm钙离子/1%胶)。
进行发酵的温度范围从25℃到35℃左右;然而要得到最佳结果,最好选用28℃到32℃。
用于培养ATCC53160菌株及产生杂多糖S-184的营养培养基的PH范围是6-8。
虽然表面培养和深层培养都可以产生S-184,但最好还是选用深层发酵。
小规模的发酵是很容易进行的,把菌株接种在一个合适的营养培养基上,然后转接到生产培养基中,在摇瓶机上,30℃恒温发酵几天。
发酵一开始是在灭过菌的含培养基的三角瓶中进行,通过1级或多级种子扩大培养。种子阶段的培养基可以组合任何合适的碳源和氮源,但是最好的碳源是葡萄糖或水解淀粉。种子瓶在30℃左右恒温室中振摇1-2天,或直到生长满意为止,一部分培养物用来接种第二级种子或直接接种到生产培养基中。如果应用中间阶段种子瓶,方法基本与上相同。也就是说,在最后一级种子瓶中取出部分培养液接种到生产培养基中。接种后的瓶在恒温振摇培养几天,培养期结束后瓶中培养物用合适的醇如异丙醇沉淀以回收其内容物。
大规模的培养,最好在带有搅拌器和能在培养基中通气的发酵缶中进行。根据这个方法,在发酵缶中配置营养培养基,加热至121℃左右灭菌。冷却后这种灭过菌的培养基利用事先已培养好生产用菌的种子来接种,发酵进行一般为2-4天。培养期间要搅拌和/或使培养基通气,并且维持温度30℃左右。这种生产S-184的方法特别别适用于大量制备。
由ATCC53160所产生的杂多糖主要由碳水化合物所组成,还含有12%-16%的蛋白质和3.0%-4.5%的酰基(以醋酸计算),碳水化合物部分包含7%-16%的糖醛酸和中性糖类:甘露糖、葡萄糖和半乳糖,它们的克分子比约为1∶3∶5左右。
下面表1给出了杂多糖S-184的组成分析的结果。
(1)乙酰基是用0.2%S-184水溶液加碱性羟胺试剂处理,随后加酸性氯化铁试剂处理而后用比色分析〔见S.He strin(1949)J.Biol.chem.180:249-261〕,氯化乙酰胆碱用作为标准。
(2)从3.1%-4.5%酰基含量(以醋酸计)是用稀碱水解氧-乙酰键后,用酶法分析来测定。
(3)S-184的中性糖是用各种方法来测定的。第一种方法是用50毫克S-184在1摩尔硫酸中100℃水介4小时;冷却后加入每毫升含3毫克的木糖溶液0.5毫升作为内标。样品中加3毫升饱和氢氧化钡溶液中和,而后加二滴刚果红和氢氧化钡一直到溶液变红为止。离心(每分钟3000转,20分钟)后取出所有样本的上清液,使蒸发干,把干样品溶于0.1毫升盐酸羟胺(每毫升40毫克)的无水吡啶溶液中,90℃加热45分钟。冷却后加入0.1毫升醋酸酐,样品再在90℃加热45分钟。糖是用它们的醛腈醋酸盐衍生物的氣-液色谱法来分离并用检定过的标样作比较进行鉴定和定量。〔J.K.Baird,M.J.Holroyde和D.C.Ellwood(1973)Carbohydr.Res.27,464-467〕。
(4)样品预处理是以样品溶于72%硫酸中,在稀释前于0℃放置1小时,并利用上面所描述的中性糖分析的第一种方法水解;结果是BD-707,BD-2117,BD-2118所得到的平均值和范围。
(5)这些样品的中性糖也可用第二种方法进行鉴别。先将2毫克左右的S-184溶于0.5摩尔的三氟醋酸中(2毫升),样品保持在100℃过夜,浓缩至干并溶于水(2毫升)中,加入25毫克硼氢化钠,2小时后溶液用Dowex 50(氢型)树脂处理以使PH下降至3.5,过滤后溶液浓缩,加甲醇(3×5毫升)一起蒸馏。残渣溶于醋酸酐(1毫升)和吡啶(1毫升)的混合液中,100℃保持1小时再浓缩。加甲苯(3×5毫升)一起蒸馏,残渣溶于二氯甲烷中用氣-液色谱法分析。得到结果是BD-707,BD-2117和BD-2118的平均值和它们的范围。
多糖中糖醛酸的百分含量是利用以下方法测定的。先用17%盐酸进行脱羧反应,将释放的二氧化碳吸收在标定好的氢氧化钠中,然后再进行回滴定〔B.L.Browning(1967)Methods of Wood Chemistry 2,632-633〕。
确定丙酮酸不存在是按下法进行即在培养管中加入1毫升含S-184每毫升2毫克的溶液再加入1毫升0.20当量盐酸,在100℃加热4小时。取0.5毫升样品水解液加入0.1毫升还源型辅酶I(NADH)和2.4毫升三乙醇胺溶液中。利用分光光度计测量吸收值的变化来计算出丙酮酸量。〔Duckworth and Yaphe,Chem,and Ind。(1970)p 747〕没有测到显著的丙酮酸量。
氮的分析是应用凯氏消化法,大约有1.9%和2.5%重量相当于11.8%-15.8%蛋白质含量。
甲基化的分析是利用S-184经过透析和冷冻干燥部分纯化的样品完成的。是根据Sandford和Conrad(1966)Biochem.5,1508-1509。所简述的步骤进行甲基化。糖类的氧-甲基醚衍生物作为艾杜糠醇醋酸盐是利用气相色谱分离到的,并经计算机
定和纯的标样对比。主要的甲基化糖经
定如下述表2所示。
表2
S-184甲基化糖残基
2,3,6三甲基六元醇(半乳糖) 1-4
2,3,6三甲基六元醇(甘露糖) 1-4
2,3,6三甲基六元醇(葡萄糖) 1-4
要了解的是虽然在这里所描述的杂多糖分析方法是得到以上所描述组成数据的实际方法,但对工艺熟悉的人来说,其他的分析方法也是可用的。应用其他分析方法,应该得到杂多糖相同的鉴别结果,然而所得到的结果在量上会稍有差异。
杂多糖S-184在水溶液中具有一些突出的性质,特别是在非常低的浓度时具有很高的粘度,很好的表面活性,很好的蛋白质可混性以及极好的稳定性/乳化性。由于这些性质,它可用作增稠,悬浮,乳化,稳定,润滑,成膜或粘合剂或作为阿拉伯胶的取代物应用于很多方面。S-184具有能在各种工业和食品中的应用是由于它的表面活性和蛋白质可混性。特别是用在以下的应用和产品中:粘合剂,墙的水泥接缝,薄浆和灰浆的保水,填泥料化合物,容器密封,锅炉防垢剂,乳胶乳化,焊条助熔剂,煤粉粘胶,陶瓷的釉和挤压,清洁剂,磨光剂,精纺丝毛,乳化液(乳胶,沥青,硅酮)银的回收,种子涂膜,防疫药或除锈剂的喷雾控制,可乳化的浓度和可流动的防疫药和除锈剂,烟草增稠,水基墨水,石印流液,皮革的抛光,水生苗和水根培养,纺织品印花和压光,制湿纸的添加剂,有助于制湿纸的保持和成型,抗滑化合物,模具的释放剂,液体树脂,粘结和包装炸药,水井钻探泥浆,石油超量和完工液,石油的促增液,化粧品,药物的悬浮和乳化。
这种胶体同样可用于食品系统如果子冻和其他高糖系统,饮料包括以柠檬酸为主的饮料,乳制品包括冰淇淋和酸乳酪,色拉的调味料,干料混合,糖衣以及釉料,糖浆,布丁,面粉的食品,缶装和瓶装食品以及烘熔的填料。
特别有价值的用途是在食品应用中,尤其如色拉的调味料,饮料和奶制品。杂多糖S-184在食品中发现特别有用是由于它的表面活性,蛋白质可混性和稳定性/乳化作用等性质。
色拉的调味剂的做法是将杂多糖S-184先在它重量的2-5倍的油中制成浆,将S-184/油的浆状物加到水中同时剧烈搅拌使之水化,再将其他的干成份混合加到水浆状物中,加西红柿浆,混合三分钟,加入含油树脂辣椒,混合三分钟,慢慢加入油,混合三分钟,加入醋,混合三分钟,用一个胶体磨使之均匀化并装瓶。
S-184作为乳化作用的稳定剂加入调味汁中,加入的量约为重量的0.1%和0.4%。已测定调味品的粘度,PH,贮存稳定性和特征性稠度结果列于下表3。
表3
在色拉调味品中的杂多糖S-184
用量 粘度*(毫泊) 稳定性(天数) 特征性稠度
% (24小时) PH 49℃(120°F) 室温 流动性
0.10 1000 3.6 7 83 光滑
薄流动
0.15 1450 3.6 31 61
0.20 1650 3.6 26 71 光滑
似乳的
超过
稳定
0.25 2700 3.6 33 365 可允许流动
半膠化
0.40 4700 3.6 未测 不允许流动
杂多糖S-184具有多种溶液特性,由于这多糖有独特的特性,而它能被辨认优于其他杂多糖。S-184有以下的多种特性:
1.粘度和切变性
A.布鲁克菲特氏
合成自来水*
合成自来水加
0.1%氯化钾
1.1.0(d每分钟60转 1080厘泊 1030厘泊
(d每分钟6转 6000厘泊
纺锤号 3
2.0.1%(UL附加器 7厘泊 7厘泊
(d每分钟6转
3.0.5%韦-布二氏
(d9.6秒-1330厘泊 320厘泊
4.1.0%去离子水(
(d每分钟60转 1460厘泊
纺锤号 3
B.切变性(韦-布二氏)
1.η(d1.92秒-14100厘泊
2.η(d9.6秒-11270厘泊
3.η(d76.8秒-1290厘泊
4.η(d384秒-1大于60厘泊(开始)
5.η(d384秒-1大于60厘泊(5分钟)
6.η(d9.6秒-11180厘泊
C.40℃贮藏(布氏)
用纺锤3号1320厘泊(d每分钟60转;
不凝结
*无离子水含1000ppm氯化钠和147ppm
氯化钙(2分子结晶水)
2.盐和染料的可混性
A.盐
1.CaCl2(饱和) 可混
2.聚磷酸铵 沉淀
3.60%NH4NO3可混
4.1%Al2(SO4)3·18H2O 可混
5.1%CaCl2·2H2O 可混
6.1%KCl 可混
B.染料
1.研磨绿 可混
2.次甲基兰 可混
杂多糖S-184可用一个未定名的产碱杆菌菌种在一个合适的营养基上发酵来制取。应用于制备这个杂多糖的微生物的生物学纯培养。按照布达佩斯条约在马里兰州洛克菲尔市,美国典型菌株收集中心保存在1985年6月19日开始,登记号为ATCC53160菌株。
A.菌落形态特征
在营养琼脂上产S-184分离菌株形成中心不透明和边缘半透明的园形菌落。产生白色到灰色色素,在30℃培养72小时后菌落直径1-3毫米。
B.细胞形态特征
细胞是革兰氏阴性,长杆状(0.8到2.3微米)带有空泡。鞭毛排列是混杂的,可看到极生,侧生和周生型式。在某些生长条件下可观察到该菌产生粉色色素。
C.生理和生化特性
生理和生化试验的结果见表4,此菌细胞色素氧化酶阳性过氧化酶阳性,属氧化代谢型。柠檬酸盐利用和硫化氢的产生阳性。它能水解淀粉,明胶,酪素和聚山梨酸酯80,USP(Difco厂生产的表面活性剂)。它们在30℃和37℃生长很好,较高温度没有进行试验。它能耐1.5%氯化钠,但不耐3%氯化钠,在PH6-10范围内均能生长。
下列为碳水化合物的产酸式样
产酸
L-阿拉伯糠
果糖
半乳糖
麦芽糖(弱)
甘露醇(弱)
甘露糖
不产酸
核糖醇
卫矛醇
乙醇
D-葡萄糖
D-木糖
肌醇
菊粉
乳糖
密二糖
α-甲基萄糖苷
棉籽糖
鼠李糖
柳醇
山梨醇
蔗糖
海藻糖
产S-184菌株能利用下列46种基质作为唯一的碳源和能源。
D-木糖
L-阿拉伯糖
D-葡萄糖
D-甘露糖
D-半乳糖
D-果糖
麦芽糖
纤维二糖
菊粉
萄糖酸盐
糖酸盐
粘酸盐
醋酸盐
丙酸盐
丁酸盐
己酸盐
庚酸盐
辛酸盐
壬酸盐
癸酸盐
丙二酸盐
琥珀酸盐
富马酸盐
L-苹果酸盐
DL-β羟丁酸盐
DL-乳酸盐
DL-甘油酸盐
柠檬酸盐
α-氧代戊二酸盐
丙酮酸盐
乙酰丙酸盐
甘露醇
甘油
D-α-丙氨酸
β-丙氨酸
L-苏氨酸
L-亮氨酸
DL-异亮氨酸
L-天冬氨酸盐
L-谷氨酸盐
L-赖氨酸
DL-鸟氨酸
L-组氨酸
L-脯氨酸
L-酪氨酸
L-苯丙氨酸
表4
S-184菌株的生理和生化试验结果
细胞色素氧化酶 +
过氧化氢酶 +
氧化发酵试验 氧化
厌气生长 -
三糖铁琼脂斜面 无变化
吲哚 -
甲基红 -
伏-普二氏试验 -
西蒙氏柠檬酸盐 +
亚硝酸盐还原 -
硝酸盐还原 -
石蕊牛奶 无变化
精氨酸脱氢酶 -
赖氨酸脱羧酶 -
鸟氨酸脱羧酶 -
苯丙氨酸脱胺酶 不生长
胨胺 -
尿素酶 -
硫化氢(从胨,胱氨酸和亚硫酸
盐培养基上) +
3-酮乳糖 -
刚果红吸收 -
磷酸酶 -
溶血(羊血) 未测
卵黄反应 ±
淀粉水解 +
明胶水解 +
酪素水解 +
七叶苷水解 -
聚山梨酸盐80水解 +
不同温度时的生长:
4℃ -
30℃ +
37℃ +
不同氯化钠浓度中的生长
1.5%(重量/体积) +
3.0%(重量/体积) -
6.5%(重量/体积) -
7.5%(重量/体积) -
10.0%(重量/体积) -
不同PH值时的生长
4 -
6 +
8 +
10 +
11 +
12 -
+=阳性 -=阴性
由于该菌是严格的好气,革兰氏阴性,并显示混杂的鞭毛型,因此根据伯奇氏细菌
定手册第八版(1974)应归属于产碱杆菌属(Alcaligenes)。
产碱杆菌属在最近出版的伯奇氏的手册(伯奇氏系统微生物学手册第一卷第一版1984年,中重新划分是不包括全部产色菌类。分离到的产S-184菌株与新划分属符合,但均不属于该手册所记载的二个种,粪产碱杆菌,反硝化产碱杆菌反硝化亚种和反硝化产碱杆菌木糖氧化亚种。因此S-184是产碱杆菌中的一个新种。
本发明的其他实施例通过这个说明书的讨论对在工艺上熟悉的人们将会是很清楚的。所以这说明书和举例被认为是只作示范,而确切的发明范围和发明构思通过所附权利要求表示出来。
例1
在营养琼脂上生长48小时的菌株接种到含酵母肉汁培养基的种子瓶,在旋转摇瓶机上30℃培养。24小时左右后,这种子接到以3%水解淀粉为碳源的E1培养基中,也在30℃旋转摇瓶机上培养。约72小时后这瓶中出现粘稠状,加入2个体积99%异丙醇得到纤维状沉淀。
E1培养基含5克磷酸氢二钾,0.1克硫酸镁,0.9克硝酸铵,0.5克Promosoy100(由Central Soya Chemurgy Division生产的酶介黄豆饼粉),30克葡萄糖,1升自来水,PH调到7.6到7.8左右。
按上述方式配制的其它酵母肉汁种子瓶,取24小时培养物接种到含不同培养基的4个瓶子中,在30℃旋转摇瓶机上培养72小时左右,然后测定其PH,粘度,胶产量和产物的粘度,其结果见下表5。
*霍氏(Hole)盐的配制是1升去离子或蒸馏水中在确加入下列组分:
H3BO3285毫克
Mncl2·4H2O 1800毫克
FeSO41360毫克
CnCl226.9毫克
Zncl220.8毫克
Cocl240.4毫克
Mg2MoO4·2H2O 25.2毫克
酒石酸钠 1770毫克
每升培养基中加1升原液。
从上述结果看到用3%葡萄糖的E1培养基是生长最好的培养基。
例2
提供大量生产杂多糖S-184的发酵步骤。
500毫升三角瓶里装100毫升酵母肉汁培养液(Difco厂),接种一环在营养琼脂平板上培养48小时的产S-184菌体细胞。在30℃每分钟400转的旋转摇瓶机上培养24小时。然后在二个500毫升三角瓶各装100毫升种子培养基中接入1%培养物。种子培养基包括:
葡萄糖 3%
K2HPO40.5%
NH4NO30.09%
MgSO4·7H2O 0.01%
酶介黄豆饼粉100 0.05%
培养基用自来水配制。这些种子瓶在30℃每分钟400转的旋转摇瓶机上培养24小时,即用作接种到含3000毫升(最终体积)同样培养基的5升发酵缶中。发酵控制在30℃,通气量每分钟1升条条件下进行。搅拌速度开始每分钟400转而后增加以保证发酵液很好的混合。24小时后约2.5升这个种子液用以接种到含20升(最后体积)培养基的30升发酵缶中。培养基成份如下:
葡萄糖 3.0%
K2HPO40.05%
NH4NO30.09%
MgSO4·7H2O 0.01%
酶介黄豆饼粉100 0.05%
Fe++1ppm
Sag5691 0.005%
(联合碳化公司生产的一种
抗泡剂)
温度保持在30℃,通气量每分钟5升,22小时后调到每分钟10升。一直保持到发酵结束。PH可用PH自动控制系统加40%氢氧化钾来调节PH使高于6.3。搅拌开始每分钟300转,22小时后增加到每分钟400转,45小时增加到每分钟700转,一直保持到发酵结束。发酵结果见下表6。
表6
发酵液 胶得量 残余碳源
缶令 PH 粘度 (克/100毫升) %
0小时 7.0 未测 未测 3.0
22小时 6.3 620厘泊 未测 未测
45小时 6.9 4200厘泊 1.26 0.41
68小时 7.2 5100厘泊 1.37 低于0.1
把发酵液加热到75℃左右,15分钟后冷却到30℃左右,然后加到三倍体积的99%异丙醇中。杂多糖呈纤维状物沉淀,容易用筛子回收。回收的纤维状物用气流干燥器在140°F干燥2.5小时,而后磨成粉。
例3
500毫升三角瓶装100毫升酵母肉汁培养液(Difco公司)接种一环营养琼脂板上培养48小时的产S-184的菌体,然后放在每分钟400转的旋转摇瓶上,在30℃培养24小时,再转接到5个装100毫升种子培养基的500毫升三角瓶中,接种量为1%。种子培养基包括:
葡萄糖 3%
K2HPO40.5%
NH4NO30.09%
MgSO4·7H2O 0.01%
酶介黄豆饼粉(Promosoy100) 0.05%
Fe 1ppm
自来水
这些瓶在每分钟400转的旋转摇瓶机上,30℃培养24小时,以用作接种到装10升(最终体积)最终培养液的14升发酵缶中的种子。发酵控制在30℃,通气量为每分钟3升,搅拌速度开始每分钟为400转,以后增加保证很好的混和。发酵培养基组分如下:
葡萄糖 3.0%
K2HPO40.05%
NH4NO30.09%
MgSO4·7H2O 0.01%
酶介黄豆饼粉(promosoy100) 0.05%
Fe++1ppm
通过自动PH控制系统加入25%氢氧化钾来控制PH大于6.5。发酵结果见下表7。
表7
发酵液 胶产量 残留物
缶令 PH 粘度 (克/100毫升) 碳源
0 7.3 未测 未测 3.15%
21 7.05 20厘泊 0.65% 1.59%
45 6.9 3450厘泊 1.22% 0.48%
78 7.1 2550厘泊 1.50% 少于0.10%
把发酵液加热到75℃左右,15分钟后冷却至30℃左右,然后加到三倍体积99%的异丙醇中,杂多糖呈纤维状沉淀,用筛子很容易回收。回收的纤维状物在气流干燥器中140°F干燥2.5小时,最后磨成粉。
勘误表
Claims (8)
1、杂多糖S-184主要由碳水化合物组成,还含有12-16%蛋白质和3.0%-4.5%酰基(以醋酸计算),碳水化合物部分包括7-16%糖醛酸,和中性糖∶甘露糖、萄葡糖和半乳糖,它们的克分子比大致为1∶3∶5。
2、一株产碱杆菌(Alcaligenes)在生物学上纯的培养物ATCC53160。
3、培养物包括由产碱杆菌(Alcaligenes)微生物组成的培养物ATCC53160,它经过通气发酵能以可回收的量产生杂多糖S-184。
4、产生杂多糖S-184的方法,包括在营养培养基中通过可同化碳源的通气发酵,使产碱杆菌ATCC53160生长,并回收所说的杂多糖S-184。
5、按照权利要求4的方法,其中可同化的碳源是碳水化合物。
6、按照权利要求5的方法,其中的碳水化合物是葡萄糖。
7、按照权利要求4的方法,其中的营养培养基实质上是无钙离子的。
8、用权利要求4的方法生产产品。
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US750,806 | 1985-06-28 |
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JP (1) | JPH0764882B2 (zh) |
KR (1) | KR890002698B1 (zh) |
CN (1) | CN1030469C (zh) |
AT (1) | ATE51646T1 (zh) |
CA (1) | CA1318622C (zh) |
DE (1) | DE3670098D1 (zh) |
DK (1) | DK305886A (zh) |
ES (1) | ES2001748A6 (zh) |
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US4269939A (en) * | 1980-06-20 | 1981-05-26 | Merck & Co., Inc. | Preparation of heteropolysaccharide S-119 |
CA1187431A (en) * | 1981-04-23 | 1985-05-21 | Suzanna M. Steenbergen | Heteropolysaccharide s-198 |
JPS5878595A (ja) * | 1981-11-05 | 1983-05-12 | Nakano Vinegar Co Ltd | 新規な酸性ヘテロ多糖類の製造法 |
-
1986
- 1986-06-19 IL IL79166A patent/IL79166A0/xx not_active IP Right Cessation
- 1986-06-23 CA CA000512201A patent/CA1318622C/en not_active Expired - Fee Related
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CN1030469C (zh) | 1995-12-06 |
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CA1318622C (en) | 1993-06-01 |
EP0211506B1 (en) | 1990-04-04 |
JPS6213401A (ja) | 1987-01-22 |
KR890002698B1 (ko) | 1989-07-24 |
ES2001748A6 (es) | 1988-06-16 |
PT82844A (en) | 1986-07-01 |
DE3670098D1 (de) | 1990-05-10 |
EP0211506A1 (en) | 1987-02-25 |
HU198220B (en) | 1989-08-28 |
ATE51646T1 (de) | 1990-04-15 |
HUT41823A (en) | 1987-05-28 |
DK305886A (da) | 1987-02-17 |
PT82844B (pt) | 1988-12-15 |
JPH0764882B2 (ja) | 1995-07-12 |
IL79166A0 (en) | 1986-09-30 |
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