CN101970640B - 解脂耶罗威亚酵母新菌株及其在生物柴油生产过程中获得的丙三醇组分的工业回收中的用途 - Google Patents
解脂耶罗威亚酵母新菌株及其在生物柴油生产过程中获得的丙三醇组分的工业回收中的用途 Download PDFInfo
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Abstract
回收由生物柴油生产产生的丙三醇组分的工业方法,用于所述方法中的解脂耶罗成亚酵母新菌株,及其用途。
Description
本发明的主题是对在生物柴油生产中获得的丙三醇组分(glycerolfraction)进行再加工的工业方法,以及特别适用于此过程的解脂耶罗威亚酵母(Yarrowia lipolytica)新菌株。
可再用的燃料油成分即所谓生物柴油的生产,主要包括由天然甘油三酯(通常为植物脂质)经转酯作用产生脂肪酸酯。美国专利2.271.619揭示了在作为催化剂的基本上无水的碱金属羟化物存在下,通过添加少于5个碳的饱和一羟基脂肪醇,将高级脂肪酸的甘油酯转化为短醇的酯的方法。依照所述专利,该过程应于86-212°F(30-100℃)下在反应器中进行。醇的量不可超过甘油酯的1.75倍,催化剂应为甘油酯质量的0.1-0.5%。
随后的专利改进或补充了此专利。美国专利2.360.844、2.383.632、2.383.580、2.383.581、2.383.614、2.383.633、2.383.596以及2.383.599,分别描述了美国2.271.619所述方法的连续变体,包括:a)向该过程中添加酸并喷雾干燥;b)增加了对于未反应的醇进行蒸馏的步骤;c)评估催化剂的活性,建议的pH为5-7;d)应用部分脂肪酸酯技术;e)回收未反应的醇,并使液体酸化,以便改善酯与丙三醇的分离;f)使用各种方法转化部分反应的甘油酯;g)与部分甲醇一起补充一羟基脂肪醇(非甲醇),以便促进液相的分离;以及h)添加溶剂以便改善相分离。
其他专利计划进行了更大范围的修改和改进。美国专利2.494.366、2.383.601、3.963.699、4.303.590、4.371.470、4.668.439、5.399.731、5.434.279以及5.525.126,也主要建立在美国2.271.619所述的工艺之上。它们分别涉及:a)向碱性催化剂中添加适量的酸性催化剂;b)进一步添加酸性的酯化催化剂;c)将该过程在恒温和从真空到大气压力的压力条件下进行,;d)添加第二次碱性催化作用阶段;e)添加第二次酯化阶段并使用吸收剂去除烷基酯;f)导入蒸汽形式的醇;g)在较低温度下增加酸的量来进行该反应;g)引入使用酸进行相分离的改进方法;以及h)使用乙 酸钙和乙酸钡的混合物作为催化剂。
不考虑所用的工艺,在甘油三酯转酯作用中形成的需要回收的主要副产物是所谓的甘油酯组分,其由可溶解的亲水性反应产物组成,意即丙三醇、未使用的催化剂残留物、脂肪酸酯和用于转酯作用过程的进一步分离阶段的其他试剂,例如磷酸和无机盐。
在生物柴油的生产中丙三醇组分的回收和/或循环成为重要的难题。
本发明的目的是,提供容易回收在生物柴油生产中出现的丙三醇组分的方法。本发明的特殊目的是,提供将丙三醇组分工业转化为容易吸收的生物质的有效方法。此方法可用于工业规模来回收丙三醇组分,并考虑了所述组分的成分因所用生物柴油生产工艺的不同而不同。所产生的生物质的特征为,高含量的易吸收蛋白质、维生素并能够用作饲料添加剂。
出乎意料的是,这些限制性难题已经在本发明中得到解决。
本发明的目的是回收由生物柴油生产产生的丙三醇组分的工业方法,特征在于将酵母解脂耶罗威亚酵母于温度25-35℃、通气量0.2-4L空气/1L培养基/分钟、维持在2.5-7.5的pH下,在由包含20.0-70.0g/l的丙三醇组分、8.5-15.0g/l的(NH4)2SO4、1.5-6.5g/l尿素、0.5-3.0g/l的MgSO4×7H2O、0.1-2.0g/l的KH2PO4和0.1-2.0g/l的酵母提取物的水性溶液组成的培养基中培养;直至培养基中的丙三醇实质性耗尽,其中以周期性方式维持该培养,在每个循环末期用部分新鲜培养基替换部分培养后液体培养基(post-culture broth)。
依据本发明的方法是指用于工业规模上,其中,优选地,该培养是维持在至少1000升的体积中。同样优选地,分离的培养后液体培养基包含15-35g/L的酵母干物质,然而干物质的产生是以1.5-3.0g/Lh的速率发生,并且生物质的产生以培养基中0.4-0.5g干生物质/g丙三醇组分的最大效率发生。优选地,干生物质的蛋白含量按质量计为30-50%。优选地,该培养利用解脂耶罗威亚酵母的A-101菌株,该菌株以登录号KP2018p保藏于IBPRS。
解脂耶罗威亚酵母的A-101菌株已经依照布达佩斯条约保藏于保藏库中,由食品和农艺工业生物技术研究所(the Institute of Biotechnology of the Food and Agriculture Industry,后文称为IBPRS,ul.Rakowiecka 36.02-532Warsaw)维持,并获得KKP 2018p登录号。其为野生型菌株,选自在设计本发明的过程中检测的该种类的许多其他菌株之中,由WroclawUniversity of Environmental and Life Sciences收集。该选择标准主要包括基于丙三醇组分的培养条件。首先,此菌株,与所检测的解脂耶罗威亚酵母的所有菌株不同,产生了生物质产生的异常良好的效率和对于有害培养条件的极大耐性,例如增加的渗透压和培养基相对低的pH。鉴于此,既然此菌株受其他微生物污染的危险如此低,培养该菌株则非常简单。同时,所产生的生物质拥有更好的营养特性,如高含量的易吸收蛋白质和维生素,尤其是维生素B。因此,它可以用作高质量的饲料添加剂。
本发明的另一个目的是以登录号KP 2018p保藏于IBPRS的解脂耶罗威亚酵母的A-101菌株。
本发明的主题还有以登录号KP 2018p保藏于IBPRS的解脂耶罗威亚酵母的A-101菌株在生物柴油生产产生的废品回收中的用途。优选地,将所产生的生物质用于饲料生产中。
实施例1
在基于来自生物柴油生产的丙三醇组分的培养基上产生酵母解脂耶罗威亚酵母生物质的过程中的培养基(培养基1)的成分,组成为(单位g/升):
丙三醇组分 20.0-70.0,优选约50.0
(NH4)2SO4- 8.5-15.0,优选约12.6
尿素 1.5-6.5,优选约4.0
MgSO4×7H2O- 0.5-3.0,优选约1.0
KH2PO4- 0.1-2.0,优选约0.5
酵母提取物 0.1-2.0,优选约0.5
自来水 至1000ml
pH- 2.5-7.5,优选约3.5-4.0
在上述比例中,称出1100L培养基1的成分,并以自来水使其达到1000L。
在培养基成分完全溶解后,将其转移至生物反应器中。向该培养基中添加100L酵母细胞,该酵母细胞是以150L工作体积在生物反应器中进行预培养,并在上述培养基中进行培养。
各种解脂耶罗威亚酵母菌株的培养条件为:
温度为25-35℃(优选约30℃±1),400-1200RPM(优选约700-800)搅拌,通气量为0.2-4L空气/L培养基/分钟(优选约1-1.5L空气/L培养基/分钟)。应当使用10N的NaOH自动维持pH。如果出现过多的泡沫,应当使用消泡剂如ACEPOL或其他消泡剂。
应当维持该培养,直至培养基中的丙三醇耗尽。应当保留200L的细胞悬浮物,应当将(培养基1)的成分加入其中使量足以至1100L,并应当加入900L的水。此培养方法(周期的、循环的)可以重复5-15次。
此酵母繁殖方案以1.5-3.0g/Lh(优选约2.5g/Lh)的速率,以至少0.4-0.5g干酵母物质/g丙三醇组分的效率(在A-101解脂耶罗威亚酵母的情况下优选约0.60g/g),产生15-35g/L(优选约33g/L)的干酵母物质。
干酵母的蛋白含量应当为30-50%,(在A-101解脂耶罗威亚酵母的情况下为42%)。
所获得的生物质可以进一步加工,例如以已知技术干燥,特别是喷雾干燥,然后作为高质量饲料添加剂分装并出售,尤其是用于牛和家禽饲料。
Claims (9)
1.回收由生物柴油生产产生的丙三醇组分的工业方法,其特征在于,将以登录号KKP 2018p保藏于IBPRS的解脂耶罗威亚酵母(Yarrowialipolyica)A-101菌株于温度25-35℃、通气量0.2-4L空气/1L培养基/分钟、维持在2.5-7.5的pH下,在由包含20.0-70.0g/l的丙三醇组分、8.5-15.0g/l的(NH4)2SO4、1.5-6.5g/l尿素、0.5-3.0g/l的MgSO4×7H2O、0.1-2.0g/l的KH2PO4和0.1-2.0g/l的酵母提取物的水性溶液组成的培养基中培养;直至所述培养基中的丙三醇实质性耗尽,其中以周期性方式维持该培养,在每个循环之后,以部分新鲜培养基替换部分培养后液体培养基。
2.如权利要求1所述的方法,其特征在于所述培养维持于至少1000升的体积中。
3.如权利要求1所述的方法,其特征在于分离的培养后液体培养基产生15-35g/L的酵母干物质。
4.如权利要求1所述的方法,其特征在于生物质的产生是以1.5-3.0g/Lh的速率进行的。
5.如权利要求1所述的方法,其特征在于所述生物质的产生是以培养基中0.4-0.5g干生物质/g丙三醇组分的总体效率发生。
6.如权利要求1所述的方法,其特征在于所述干生物质的蛋白含量按质量计为30-50%。
7.以登录号KKP 2018 p保藏于IBPRS的解脂耶罗威亚酵母(Yarrowia lipolytica)的A-101菌株。
8.以登录号KKP 2018 p保藏于IBPRS的解脂耶罗威亚酵母(Yarrowia lipolyica)的A-101菌株在生物柴油生产的废品回收中的用途。
9.如权利要求8所述的用途,其特征在于将所产生的生物质用于饲料生产中。
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PCT/PL2008/000077 WO2009061225A1 (en) | 2007-11-05 | 2008-11-05 | A new strain of yarrowia lipolytica and its use in the industrial reclamation of glycerol fractions obtained during biodiesel production |
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US3843800A (en) * | 1968-11-08 | 1974-10-22 | Arend Langejan | Preparation of active dried baker's yeast |
US3713976A (en) * | 1971-06-21 | 1973-01-30 | Gulf Research Development Co | Cultivation of micro-organisms on hydrocarbons |
MX348214B (es) * | 2006-03-10 | 2017-06-05 | Archer Daniels Midland Co | Metodos y composiciones para productividad aumentada en animales. |
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2008
- 2008-11-05 EP EP08847410.1A patent/EP2231855B1/en active Active
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WO2007017356A1 (fr) * | 2005-08-09 | 2007-02-15 | Specialites Pet Food | Facteur d'appetence fermente et depourvu de proteines animales pour animaux |
Non-Patent Citations (3)
Title |
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IMANDI S.B..Optimization of medium constituents for the production of citric acid.《ENZYME AND MICROBIAL TECHNOLOGY》.2007,第40卷(第5期),1367-1372. * |
PANPANIKOLAOU S..Lipid production by Yarrowia lipolytica growing on industrial glycerol in a single-stage continuous culture.《BIORESOURCE TECHNOLOGY》.2002,第82卷(第1期),43-49. * |
PANPANIKOLAOU S..Yarrowia lipolytica as a potential producer of citric acid.《JOURNAL OF APPLIED MICROBIOLOGY》.2002,第92卷(第1期),737-744. * |
Also Published As
Publication number | Publication date |
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WO2009061225A1 (en) | 2009-05-14 |
EP2231855B1 (en) | 2015-10-07 |
PL383685A1 (pl) | 2009-05-11 |
EP2231855A1 (en) | 2010-09-29 |
CN101970640A (zh) | 2011-02-09 |
US20100317076A1 (en) | 2010-12-16 |
PL215829B1 (pl) | 2014-01-31 |
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