WO2019056919A1 - Microbial oil containing low chloropropanol and preparation method thereof - Google Patents
Microbial oil containing low chloropropanol and preparation method thereof Download PDFInfo
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- the fermentation temperature is maintained at 25-35 ° C, and the pH is maintained at 6-8.
- the culture medium needs to contain nutrients such as nitrogen, phosphorus, and potassium.
- the fermentation broth is subjected to a bastard treatment; the temperature of the pasteurization treatment is 70-90 ° C, preferably 80 ° C.
- the fermentation of the fermentation broth at a certain temperature can inactivate the lipase in the fermentation broth and further enhance the inhibitory effect on lipase activity.
- the combination of lipase inhibitor and bark process can effectively inhibit or reduce the decomposition of triglyceride in microbial oil, reduce the content of glycidyl ester or chloropropanol in microbial oil, and increase glycerol The content of fat.
- microchlorohydrin having a low chloropropanol content prepared by the above method.
- the components of the lipase inhibitor are obtained by the method of fermentation by plants, which are non-toxic substances, do not adversely affect the environment, and do not have toxicological effects on the final microbial oil;
- Microbial oil was prepared in the same manner as in Example 10 except that no lipase inhibitor was added.
- the contents of the main components in the microbial oil and fat are shown in Table 10.
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Abstract
The present invention provides a microbial oil containing low chloropropanol and a preparation method thereof, comprising: adding lipase inhibitor to a fermentation liquid of microbial fermentation, then extracting, refining and deodorizing to reduce the content of glycidyl ester and/or chloropropanol in the obtained microbial oil. The lipase inhibitor is capable of inhibiting the activity of lipases in microbial cells in the fermentation liquid.
Description
交叉引用cross reference
本申请要求2017年9月25日提交的专利名称为“一种低氯丙醇微生物油脂及其制备方法”的第201710874273.2号中国专利申请的优先权，其全部公开内容通过引用整体并入本文。The present application claims the benefit of priority to the benefit of the benefit of the benefit of the benefit of the benefit of the benefit of the benefit of the benefit of the benefit of the benefit of the benefit of the benefit of the benefit of the benefit of the benefit of the benefit of the benefit of the benefit of the benefit of the benefit of the disclosure of the entire disclosure of
本发明涉及微生物油脂制备技术，更具体地，涉及一种低氯丙醇微生物油脂及其制备方法。The invention relates to a microbial oil preparation technology, and more particularly to a low chloropropanol microbial oil and a preparation method thereof.
氯丙醇是丙三醇中羟基被氯取代后形成的一类化合物的总称，根据氯原子取代的个数及位置不同,可将氯丙醇分为单氯丙二醇和双氯丙醇两类。其中，3-氯-l,2-丙二醇(3-MCPD)具有干扰人体内性激素平衡，使***数量减少从而使雄性动物生殖能力减弱的作用。此外，还具有慢性毒性、致癌性和神经毒性。Chloropropanol is a general term for a class of compounds in which hydroxy groups in glycerol are substituted by chlorine. Chloropropanol can be classified into two types: monochloropropanediol and dichloropropanol depending on the number and position of chlorine atoms. Among them, 3-chloro-l,2-propanediol (3-MCPD) has the effect of disturbing the balance of sex hormones in the human body, reducing the number of sperm and reducing the reproductive capacity of male animals. In addition, it has chronic toxicity, carcinogenicity and neurotoxicity.
微生物油脂是采用酵母、霉菌、细菌或藻类等微生物在一定条件下，利用碳水化合物、碳氢化合物或普通油脂为碳源、氮源，以此制备微生物油脂。由于此种制备方法具有油脂含量高、生产周期短、不受季节影响、不占用耕地等诸多优点，其应用的领域较为广泛，也应用以食品领域。Microbial oils and fats are prepared by using microorganisms such as yeast, mold, bacteria or algae under certain conditions, using carbohydrates, hydrocarbons or common fats and oils as carbon sources and nitrogen sources. Because of the advantages of high oil content, short production cycle, no seasonal influence, and no occupation of cultivated land, the preparation method has wide application fields and is also applied to the food field.
在制备微生物油脂的过程中，由于缩水甘油酯(GEs)是缩水甘油和脂肪酸的酯化产物，它存在环氧基结构，属于一类末端环氧酯，环氧基的化学性质活泼，可与细胞内亲核物质反应。因此，在制备过程中或后续作为食品的食用过程中，不可避免会产生缩水甘油酯，而缩水甘油酯极易水解而形成氯丙醇。同时，由于缩水甘油酯的产生，在制备过程中的加热过程会加快3-MCPD的生成反应，而使最终微生物油脂的3-MCPD的含量较高。In the process of preparing microbial oil, since glycidyl ester (GEs) is an esterification product of glycidol and fatty acid, it has an epoxy structure and belongs to a kind of terminal epoxy ester. The epoxy group is chemically active and can be used. Intracellular nucleophile reaction. Therefore, in the course of preparation or subsequent consumption as a food, glycidyl ester is inevitably produced, and glycidyl ester is easily hydrolyzed to form chloropropanol. At the same time, due to the production of glycidyl ester, the heating process during the preparation process accelerates the formation reaction of 3-MCPD, and the content of 3-MCPD of the final microbial oil is higher.
发明内容Summary of the invention
本发明提供一种克服上述问题或者至少部分地解决上述问题的低氯丙醇微生物油脂及其制备方法，以解决食用微生物油脂中缩水甘油酯或氯丙醇含量高的技术问题。The present invention provides a low chloropropanol microbial oil and fat which overcomes the above problems or at least partially solves the above problems, and a preparation method thereof, to solve the technical problem of high content of glycidyl ester or chloropropanol in edible microbial oil.
根据本发明的一个方面，提供一种低氯丙醇微生物油脂，该微生物油脂中，氯丙醇含量小于0.1ppm、TAG含量≥95％、DHA、ARA或EPA含量≥40％。According to an aspect of the present invention, there is provided a low chloropropanol microbial oil having a chloropropanol content of less than 0.1 ppm, an TAG content of ≥95%, and a DHA, ARA or EPA content of ≥40%.
根据本发明的另一个方面，提供一种低氯丙醇微生物油脂的制备方法，其在微生物菌种发酵的发酵液中添加脂肪酶抑制剂，以抑制发酵液中微生物细胞内脂肪酶的活性，降低脂肪酶的催化合成作用。微生物菌种发酵后的产物再依次经提取、精炼和脱臭处理后，得到微生物油脂。在所得的微生物油脂中，其缩水甘油酯和/或氯丙醇的含量大大降低，但并不会影响到微生物油脂的油脂产出率。According to another aspect of the present invention, a method for preparing a microchloropropanol microbial oil and fat is provided, which comprises adding a lipase inhibitor to a fermentation broth of a microbial strain to inhibit the activity of a lipase in a microbial cell in the fermentation broth, Reduce the catalytic synthesis of lipase. The product after fermentation of the microbial strain is subjected to extraction, refining and deodorization treatment in order to obtain microbial oil. In the obtained microbial oil, the content of glycidyl ester and/or chloropropanol is greatly reduced, but does not affect the fat yield of the microbial oil.
具体地，在微生物菌种的发酵过程中，为使微生物菌种在培养基中的发酵效果更好，其发酵的温度保持在25-35℃，pH保持在6-8。具体地，培养基需含有氮、磷、钾等营养元素。Specifically, in the fermentation process of the microbial strain, in order to make the fermentation effect of the microbial strain in the medium better, the fermentation temperature is maintained at 25-35 ° C, and the pH is maintained at 6-8. Specifically, the culture medium needs to contain nutrients such as nitrogen, phosphorus, and potassium.
具体地，在微生物菌种发酵得到的发酵液中添加脂肪酶抑制剂，该脂肪酶抑制剂能够有效的抑制微生物细胞内脂肪酶的活性。在应用微生物菌种以提炼微生物油脂的过程中，由于在目标油脂合成达到饱和的情况下，微生物细胞内的脂肪酶能够催化合成大量的缩水甘油酯，进而使最终的微生物油脂中的氯丙醇含量较高，影响微生物油脂的品质。Specifically, a lipase inhibitor is added to the fermentation broth obtained by fermentation of the microorganism species, and the lipase inhibitor can effectively inhibit the activity of the lipase in the microbial cells. In the process of applying microbial strains to refine microbial oils, the lipase in the microbial cells can catalyze the synthesis of a large amount of glycidyl ester in the process of saturation of the target oil and fat, thereby making the chloropropanol in the final microbial oil. High content, affecting the quality of microbial oils.
因此，通过在微生物菌种提炼微生物油脂的初始阶段加入脂肪酶抑制剂，影响微生物细胞内脂肪酶的活性，以降低最终产物中的氯丙醇含量。即在微生物油脂制备的初始阶段，即对缩水甘油酯的含量进行控制，进而有效降低其在后续的反应中或食用过程中产生氯丙醇的几率。在添加脂肪酶抑制剂的过程中，无需对微生物油脂制备过程进行适应性调整，也无需设置其他额外处理过程。Therefore, by adding a lipase inhibitor in the initial stage of refining the microbial oil in the microbial strain, the activity of the lipase in the microbial cells is affected to reduce the chloropropanol content in the final product. That is, in the initial stage of preparation of the microbial oil, the content of the glycidyl ester is controlled, thereby effectively reducing the probability of producing chloropropanol in the subsequent reaction or in the edible process. In the process of adding a lipase inhibitor, there is no need to adapt the microbial oil preparation process or to set up additional processing.
在另一个具体的实施例中，所述脂肪酶抑制剂为：木犀草素、橙皮苷、 新橙皮苷、枸橘苷、齐墩果酸中的一种或几种；优选为木犀草素。例如，木犀草素可以是由水藻叶发酵后进行分离得到的有效成分。In another specific embodiment, the lipase inhibitor is one or more of luteolin, hesperidin, neohesperidin, quercetin, oleanolic acid; preferably sylvestris Prime. For example, luteolin may be an active ingredient obtained by fermentation of algae leaves.
具体地，脂肪酶抑制剂由植物进行发酵及相应的分离处理后所得的有效成分，均为无毒的物质，且成本低廉。在其处理、获取和使用过程中，不会对环境造成不利影响，也不会对微生物菌种产生不良的毒理影响。同时，脂肪酶抑制剂由植物本体发酵的过程与微生物菌种的发酵过程相似，因此，在使用脂肪酶抑制剂抑制发酵液中微生物细胞内脂肪酶的活性时，并不会对微生物菌种发酵和后续处理过程产生不良影响，也不会影响微生物油脂的油脂产出率。Specifically, the active ingredients obtained by the fermentation of the lipase inhibitor and the corresponding separation treatment are all non-toxic substances, and the cost is low. During its handling, acquisition and use, it will not adversely affect the environment and will not cause adverse toxicological effects on microbial species. At the same time, the process of fermentation of the lipase inhibitor by the plant body is similar to the fermentation process of the microbial strain. Therefore, when the lipase inhibitor is used to inhibit the activity of the lipase in the microbial cell in the fermentation broth, the microbial strain is not fermented. And the subsequent treatment process has an adverse effect, and will not affect the oil production rate of microbial oil.
在另一个具体的实施例中，在所述微生物菌种发酵的过程中，所述发酵液中的含油率达到4.0％-7.0％时，添加所述脂肪酶抑制剂；优选地，在发酵液中的含油率达到5.0％-6.0％时添加脂肪酶抑制剂。In another specific embodiment, during the fermentation of the microbial strain, when the oil content in the fermentation broth reaches 4.0%-7.0%, the lipase inhibitor is added; preferably, in the fermentation broth A lipase inhibitor is added when the oil content in the medium reaches 5.0% to 6.0%.
具体地，在微生物菌种发酵过程中，每隔一段时间即对发酵液进行取样，或按照发酵程度的不同合理安排取样时间，并对其中的各项相关指标进行检测，以监控发酵的程度。各项相关指标可以是发酵液中的菌体的油脂含量，以及发酵液中总糖、还原糖、氨基氮或菌体浓度。Specifically, in the fermentation process of the microbial strain, the fermentation liquid is sampled at intervals, or the sampling time is reasonably arranged according to the degree of fermentation, and various related indicators are detected to monitor the degree of fermentation. The relevant indicators may be the oil content of the bacteria in the fermentation broth, and the total sugar, reducing sugar, amino nitrogen or bacterial concentration in the fermentation broth.
在另一个具体的实施例中，在所述微生物菌种发酵完成后、在对所述发酵液细胞进行破壁处理开始的0-1h内添加所述脂肪酶抑制剂；优选为，在所述破壁处理开始时立即添加脂肪酶抑制剂，即在所述微生物细胞发酵过程结束后、开始对发酵液中的细胞进行破壁处理的同时，立即添加脂肪酶抑制剂。In another specific embodiment, after the fermentation of the microbial strain is completed, the lipase inhibitor is added within 0-1 h of the start of the cell wall treatment of the fermentation broth; preferably, in the Immediately after the breakage treatment, a lipase inhibitor is added, that is, a lipase inhibitor is added immediately after the end of the fermentation process of the microbial cell, and the cell in the fermentation broth is started to be broken.
具体地，微生物菌种发酵完成后，即开始对发酵液中的微生物细胞进行破壁处理。破壁处理可采用生物酶解法或机械破壁，例如向发酵完成后的发酵液中加入碱性蛋白酶进行破壁处理。在发酵过程完成后，可将破壁处理时的发酵液的pH调节至7-10的范围内，并通过显微镜镜检，以检测发酵液中微生物细胞的破壁效果，破壁至没有完整的细胞为止。Specifically, after the fermentation of the microbial strain is completed, the microbial cells in the fermentation broth are subjected to a wall breaking treatment. The wall breaking treatment may be carried out by bio-enzymatic method or mechanical breaking, for example, by adding an alkaline protease to the fermentation liquid after the fermentation is completed, and breaking the wall. After the fermentation process is completed, the pH of the fermentation broth during the wall breaking treatment can be adjusted to a range of 7-10, and microscopic examination is performed to detect the breaking effect of the microbial cells in the fermentation broth, and the wall is broken until it is intact. So far.
在微生物菌种发酵末期，或在破壁处理的初始阶段，发酵液中微生物 细胞内脂肪酶的催化合成作用加剧，在目标油脂合成接近饱和的情况下，氯丙醇的合成几率会大幅上升。At the end of the fermentation of microbial strains, or in the initial stage of the wall-breaking treatment, the catalytic synthesis of lipase in the microbial cells in the fermentation broth is intensified. When the target oil synthesis is nearly saturated, the synthesis probability of chloropropanol will increase significantly.
具体地，由于所添加的脂肪酶抑制剂能够抑制微生物细胞内脂肪酶的催化合成活性。当在发酵的初始阶段添加脂肪酶抑制剂时，会减弱微生物细胞内脂肪酶的活性，降低发酵的效率或品质，从而降低微生物菌种的油脂产出效率。Specifically, since the added lipase inhibitor is capable of inhibiting the catalytic synthesis activity of the lipase in the microbial cells. When a lipase inhibitor is added in the initial stage of fermentation, the activity of the lipase in the microbial cells is weakened, and the efficiency or quality of the fermentation is lowered, thereby reducing the oil production efficiency of the microbial strain.
当在破壁处理的后期添加脂肪酶抑制剂时，由于发酵过程完成后，发酵液中的目标油脂的含量处于峰值或接近峰值的状态，微生物细胞内脂肪酶的催化合成活性较高，在破壁处理过程中即会形成大量的缩水甘油酯，使后续产物中的氯丙醇含量高，即在破壁后期，大量的氯丙醇或易于分解为氯丙醇的前体已经生成。因此，在破壁后期添加脂肪酶抑制剂的效果不佳。When a lipase inhibitor is added in the late stage of the wall breaking treatment, since the content of the target oil in the fermentation broth is at or near the peak after the completion of the fermentation process, the catalytic activity of the lipase in the microbial cell is high, and is broken. A large amount of glycidyl ester is formed during the wall treatment, so that the chloropropanol content in the subsequent product is high, that is, in the late stage of the broken wall, a large amount of chloropropanol or a precursor which is easily decomposed into chloropropanol has been formed. Therefore, the effect of adding a lipase inhibitor at the end of the broken wall is not good.
因此，在微生物菌种发酵末期，或在破壁处理的初始阶段，即发酵液中的含油率接近峰值的期间加入脂肪酶抑制剂，以抑制发酵液中微生物细胞内脂肪酶的活性，降低在其催化作用下合成的缩水甘油酯含量，从而使得最终产物中的氯丙醇含量降低。Therefore, at the end of the fermentation of the microbial strain, or during the initial stage of the broken wall treatment, that is, the oil content in the fermentation broth is close to the peak period, a lipase inhibitor is added to inhibit the activity of the lipase in the microbial cells in the fermentation broth, and The glycidyl ester content synthesized under the catalysis causes a decrease in the chloropropanol content in the final product.
在另一个具体的实施例中，所述脂肪酶抑制剂的添加量与所述发酵液的质量比为0.005％-0.05％，优选为0.02％-0.03％。具体地，在发酵液中添加的脂肪酶抑制剂的质量与发酵液的质量比保持在合适的范围，能够提高所添加的脂肪酶抑制剂对发酵液中微生物细胞内脂肪酶活性的抑制作用。In another specific embodiment, the mass ratio of the lipase inhibitor to the fermentation broth is from 0.005% to 0.05%, preferably from 0.02% to 0.03%. Specifically, the mass ratio of the lipase inhibitor added to the fermentation broth to the fermentation broth is maintained in an appropriate range, and the inhibitory effect of the added lipase inhibitor on the lipase activity in the microbial cells in the fermentation broth can be enhanced.
具体地，由于在发酵的末期或破壁处理的初始阶段，发酵液中的含油率较高，即目标油脂的含量较高，在此情形下，脂肪酶催化合成缩水甘油酯的效率提高。在脂肪酶抑制剂的作用下，能够较低脂肪酶的催化合成效率。但是，添加的脂肪酶抑制剂含量过高，一方面增加了生产成本，另一方面也会抑制其他酶类，如碱性蛋白酶的活性。Specifically, since the oil content in the fermentation liquid is high at the end stage of the fermentation or in the initial stage of the wall breaking treatment, that is, the content of the target oil is high, in this case, the efficiency of the lipase-catalyzed synthesis of the glycidyl ester is improved. Under the action of lipase inhibitors, the catalytic synthesis efficiency of lower lipase can be lowered. However, the excessive addition of lipase inhibitors increases the production cost on the one hand and inhibits the activity of other enzymes such as alkaline protease on the other hand.
在另一个具体的实施例中，在微生物菌种发酵完成后，对发酵液进行巴杀处理；所述巴杀处理的温度为70-90℃，优选为80℃。在一定温度下 对发酵液进行巴杀处理，能够使发酵液中的脂肪酶失活，进一步增强对脂肪酶活性的抑制效果。采用脂肪酶抑制剂与巴杀工艺处理结合的处理方式，能够有效地抑制或减弱微生物油脂中甘油三脂的分解，降低微生物油脂中缩水甘油酯或氯丙醇的含量，同时，能够增加甘油三脂的含量。In another specific embodiment, after the fermentation of the microbial strain is completed, the fermentation broth is subjected to a bastard treatment; the temperature of the pasteurization treatment is 70-90 ° C, preferably 80 ° C. The fermentation of the fermentation broth at a certain temperature can inactivate the lipase in the fermentation broth and further enhance the inhibitory effect on lipase activity. The combination of lipase inhibitor and bark process can effectively inhibit or reduce the decomposition of triglyceride in microbial oil, reduce the content of glycidyl ester or chloropropanol in microbial oil, and increase glycerol The content of fat.
在另一个具体的实施例中，所述微生物菌种包括：酵母、裂殖壶菌、双鞭甲藻、微球藻、破囊壶菌或高山被孢霉。In another specific embodiment, the microbial species comprises: yeast, Schizochytrium, dinoflagellate, micrococcus, thraustochytrid or Mortierella alpina.
在另一个具体的实施例中，向所述发酵液经过过滤、烘干处理所得的菌体中加入有机溶剂，然后升温至30-45℃，再经剪切、搅拌、过滤和脱溶后得到微生物毛油。具体地，当采用有机溶剂为己烷或丁烷，其升温后的温度为30-45℃。具体地，此过程属于微生物油脂的提取过程，即发酵液经过滤、烘干，加入有机溶剂后，再升温，并经剪切、搅拌和脱溶的过程，是提取微生物毛油的过程，属于获取微生物油脂的提取过程。In another specific embodiment, the organic solvent is added to the cells obtained by the filtration and drying treatment of the fermentation broth, and then the temperature is raised to 30-45 ° C, and then obtained by shearing, stirring, filtering and desolvation. Microbial hair oil. Specifically, when the organic solvent is hexane or butane, the temperature after the temperature rise is 30 to 45 °C. Specifically, the process belongs to the process of extracting microbial oil, that is, the process of extracting microbial oil by filtering, drying, adding organic solvent, heating, and shearing, stirring and desolvating. Obtain the extraction process of microbial oils.
在另一个具体的实施例中，将所述发酵液经过离心机浓缩得到浓缩菌体，浓缩菌体升温至55-65℃、调节pH至8.0-10.0、加入碱性蛋白酶酶解破壁后在线升温75-85℃，并直接进行离心分离处理，以得到微生物毛油。In another specific embodiment, the fermentation broth is concentrated by a centrifuge to obtain a concentrated bacterial cell, and the concentrated bacteria are heated to 55-65 ° C, adjusted to a pH of 8.0-10.0, and added to the alkaline protease to break the wall and then online. The temperature was raised at 75-85 ° C and directly subjected to centrifugation to obtain microbial hair oil.
具体地，对经过破壁处理的菌体直接升温处理后，无需再向菌体中添加有机溶剂，直接进行离心分离，即可达到提取的目的。采用升温后离心处理的方法，能够简化处理过程，节约溶剂回收成本，减少环境污染。Specifically, after directly heating the cells subjected to the broken wall treatment, it is not necessary to add an organic solvent to the cells, and the centrifugation is directly performed to achieve the purpose of extraction. The method of centrifugal treatment after heating can simplify the process, save solvent recovery cost, and reduce environmental pollution.
制备所得的微生物毛油再依次经精炼和脱臭处理，以获取符合质量要求的微生物油脂。具体地，精炼过程包括脱胶、碱炼和脱色处理，上述脱胶、碱炼、脱色、脱臭处理过程采用常规的处理工艺进行处理即可。The obtained microbial oil is then subjected to refining and deodorization treatment in order to obtain microbial oils meeting the quality requirements. Specifically, the refining process includes degumming, alkali refining, and decolorization treatment, and the above degumming, alkali refining, decolorizing, and deodorizing treatment processes may be performed by a conventional treatment process.
在另一个具体的实施例中，在对所得的微生物毛油依次经脱胶、碱炼、脱色和脱臭处理后，再进一步对所得产物采用分离蒸馏的方法进行处理，以进一步降低所得微生物油脂中氯丙醇的含量。In another specific embodiment, after the obtained microbial oil is sequentially subjected to degumming, alkali refining, decolorization and deodorization treatment, the obtained product is further subjected to separation and distillation to further reduce chlorine in the obtained microbial oil. The content of propanol.
具体地，采用分子蒸馏处理时的真空度为0-0.5Pa，优选为0.1-0.2Pa。具体地，在此真空度范围内，能够使氯丙醇分子与微生物油脂中的有效成分充分的分离，使自由程较大的氯丙醇分子从微生物油脂中分离出去，以 进一步降低所得微生物油脂中的氯丙醇含量。Specifically, the degree of vacuum when the molecular distillation treatment is employed is 0 to 0.5 Pa, preferably 0.1 to 0.2 Pa. Specifically, in the vacuum range, the chloropropanol molecules can be sufficiently separated from the active ingredients in the microbial oil, and the free-range chloropropanol molecules can be separated from the microbial oil to further reduce the obtained microbial oil. The chloropropanol content.
在另一个具体的实施例中，分子蒸馏的温度保持在150-200℃，优选为180-190℃。具体地，分子蒸馏的温度范围保持在此范围内，能够使异丙醇处于气体状态，同时，利用氯丙醇与微生物油脂中有效组分的自由程的不同而更有效的分离氯丙醇，降低微生物油脂中的氯丙醇含量。In another specific embodiment, the temperature of the molecular distillation is maintained at 150-200 ° C, preferably 180-190 ° C. Specifically, the temperature range of the molecular distillation is kept within this range, and the isopropanol can be in a gaseous state, and at the same time, the chloropropanol is more effectively separated by using the difference in the free path of the effective component of the chloropropanol and the microbial oil. Reduce the chloropropanol content of microbial oils and fats.
在另一个具体的实施例中，向微生物菌种发酵的发酵液中添加脂肪酶抑制剂，并对完成发酵后的发酵液进行巴杀处理；In another specific embodiment, a lipase inhibitor is added to the fermentation broth fermented by the microbial strain, and the fermentation broth after the fermentation is completed;
其中，所述发酵液中的含油率达到5％-6％时，添加脂肪酶抑制剂；所述脂肪酶抑制剂为木犀草素或齐墩果酸；所述脂肪酶抑制剂的添加量与所述发酵液的质量比为0.01％-0.03％；Wherein, when the oil content in the fermentation liquid reaches 5%-6%, a lipase inhibitor is added; the lipase inhibitor is luteolin or oleanolic acid; the amount of the lipase inhibitor is added The mass ratio of the fermentation broth is 0.01%-0.03%;
所述巴杀处理的温度为80℃；The temperature of the paste treatment is 80 ° C;
所述微生物菌种为裂殖壶藻或高山被包霉；The microbial strain is a schistosomiasis or a mountain is covered with mold;
进一步地，所述发酵液经提炼、精炼、脱臭处理后，再经分子蒸馏处理，所述分子蒸馏处理的真空度为0.2-0.5Pa、处理温度为150-200℃。Further, the fermentation liquid is subjected to refining, refining, deodorization treatment, and then subjected to molecular distillation treatment, and the molecular distillation treatment has a vacuum degree of 0.2-0.5 Pa and a treatment temperature of 150-200 ° C.
可以理解的是，采用不同的微生物菌种，该制备方法可用于制备多种微生物油脂，例如二十碳四烯酸、二十二碳六烯酸、二十碳五烯酸等。It will be appreciated that the use of different microbial species can be used to prepare a variety of microbial oils, such as arachidonic acid, docosahexaenoic acid, eicosapentaenoic acid, and the like.
根据本发明的另一个方面，还提供一种由上述方法制备的低氯丙醇含量的微生物油脂。According to another aspect of the present invention, there is also provided a microchlorohydrin having a low chloropropanol content prepared by the above method.
本发明的有益效果主要如下：The beneficial effects of the present invention are mainly as follows:
(1)在微生物菌种发酵的发酵液中添加能够抑制发酵液中微生物细胞内脂肪酶的活性，从而降低其催化合成缩水甘油酯的几率，进而降低最终所得的微生物油脂中缩水甘油酯和/或氯丙醇的含量；并且，其适用性好，无需对已有的工艺过程进行特殊的适应性改进；(1) Adding to the fermentation broth fermented by the microbial strain can inhibit the activity of the lipase in the microbial cells in the fermentation broth, thereby reducing the probability of catalytically synthesizing the glycidyl ester, thereby reducing the glycidyl ester in the finally obtained microbial oil and/or Or the content of chloropropanol; and, its applicability is good, no special adaptation to the existing process is required;
(2)脂肪酶抑制剂的成分是由植物通过发酵的方法提取而得，均为无毒物质，不会对环境有不利影响，也不会对最终的微生物油脂有产生毒理作用；(2) The components of the lipase inhibitor are obtained by the method of fermentation by plants, which are non-toxic substances, do not adversely affect the environment, and do not have toxicological effects on the final microbial oil;
(3)在微生物菌种的发酵末期或破壁处理初期添加脂肪酶抑制剂， 能够保证微生物菌种的油脂产出效率不受影响，同时，也能够降低最终微生物油脂中的缩水甘油酯和/或氯丙醇含量；(3) Adding a lipase inhibitor at the end of the fermentation of the microbial strain or at the beginning of the broken wall treatment can ensure that the oil production efficiency of the microbial strain is not affected, and at the same time, the glycidyl ester in the final microbial oil can be reduced and/or Or chloropropanol content;
(4)在微生物发酵完成后，对发酵液进行巴杀处理，能够进一步移植脂肪酶的活性，降低微生物油脂中氯丙醇含量；(4) After the completion of the microbial fermentation, the fermentation broth is subjected to a pasteurization treatment, and the lipase activity can be further transplanted to reduce the chloropropanol content in the microbial oil;
(5)对微生物油脂进行分子蒸馏处理，能够直接降低油脂中氯丙醇含量。(5) The molecular distillation treatment of the microbial oil and fat can directly reduce the chloropropanol content in the oil and fat.
下面结合实施例，对本发明的具体实施方式作进一步详细描述。以下实施例用于说明本发明，但不用来限制本发明的范围。The specific embodiments of the present invention are further described in detail below with reference to the embodiments. The following examples are intended to illustrate the invention but are not intended to limit the scope of the invention.
实施例1Example 1
以高山被包霉作为发酵菌种，在发酵所得发酵液中的含油率为5.31％时，加入发酵液质量0.025％的木犀草素，发酵液经板框过滤得到浓缩菌体，浓缩菌体烘干后，加入己烷并升温至80℃，剪切搅拌，筛网过滤脱溶后得到二十碳四烯酸(ARA)毛油。该毛油经酸法脱胶、碱炼、脱色、脱臭后产出微生物油脂。该微生物油脂再进一步在0.2Pa、185℃条件下，经分子蒸馏处理得到精制微生物油脂。所得产品中的主要成分见表1。When the oil content in the fermentation broth obtained from the fermentation is 5.31%, the luteolin with a mass of 0.025% of the fermentation broth is added, and the fermentation broth is filtered through the plate frame to obtain the concentrated bacteria, and the concentrated bacteria are dried. After drying, hexane was added and the temperature was raised to 80 ° C, sheared and stirred, and the mesh was filtered and desolvated to obtain an eicosatetraenoic acid (ARA) hair oil. The oil is degummed by acid method, alkali refining, decolorizing, and deodorizing to produce microbial oil. The microbial oil and fat is further subjected to molecular distillation treatment at 0.2 Pa and 185 ° C to obtain a purified microbial fat. The main components in the obtained product are shown in Table 1.
表1 微生物油脂主要指标对比Table 1 Comparison of main indicators of microbial oil
实施例2Example 2
以高山被包霉作为发酵菌种，在发酵所得发酵液中的含油率为4.53％时，加入发酵液质量0.01％的齐墩果酸，发酵液经板框过滤得到浓缩菌体，浓缩菌体烘干后，加入己烷并升温至80℃，剪切搅拌，筛网过滤脱溶得到二十碳四烯酸(ARA)毛油。该毛油经酸法脱胶、碱炼、脱色、脱臭后产出微生物油脂，该微生物油脂再进一步在0.5Pa、195℃条件下，经分 子蒸馏处理得到精制微生物油脂。所得微生物油脂中的主要成分见表2。When the oil content of the fermentation broth obtained by fermentation is 4.53%, the oleanolic acid of 0.01% by mass of the fermentation broth is added, and the fermentation broth is filtered through the plate frame to obtain the concentrated bacteria, and the concentrated bacteria are concentrated. After drying, hexane was added and the temperature was raised to 80 ° C, sheared and stirred, and screen-filtered and desolvated to obtain arachidonic acid (ARA) hair oil. The oil is subjected to acid degumming, alkali refining, decolorization, and deodorization to produce microbial oil, and the microbial oil is further subjected to molecular distillation at 0.5 Pa and 195 ° C to obtain a purified microbial oil. The main components in the obtained microbial oil and fat are shown in Table 2.
表2 微生物油脂主要指标对比Table 2 Comparison of main indicators of microbial oil
实施例3Example 3
以高山被包霉作为发酵菌种，在发酵所得发酵液中的含油率为6.27％时，加入发酵液质量0.01％的齐墩果酸，发酵液经板框过滤得到浓缩菌体，浓缩菌体烘干后，加入己烷并升温至80℃，剪切搅拌，筛网过滤脱溶得到二十碳四烯酸(ARA)毛油。该毛油经酸法脱胶、碱炼、脱色、脱臭后产出微生物油脂，该微生物油脂再进一步在0.4Pa、150℃条件下，经分子蒸馏处理得到精制微生物油脂。所得微生物油脂中的主要成分见表3。When the oil content of the fermentation broth obtained by fermentation is 6.27%, the oleanolic acid of 0.01% by mass of the fermentation broth is added, and the fermentation broth is filtered through the plate frame to obtain the concentrated bacteria, and the concentrated bacteria are concentrated. After drying, hexane was added and the temperature was raised to 80 ° C, sheared and stirred, and screen-filtered and desolvated to obtain arachidonic acid (ARA) hair oil. The oil is subjected to acid degumming, alkali refining, decolorization, and deodorization to produce microbial oil, and the microbial oil is further subjected to molecular distillation treatment at 0.4 Pa and 150 ° C to obtain a refined microbial oil. The main components in the obtained microbial oil and fat are shown in Table 3.
表3 微生物油脂主要指标对比Table 3 Comparison of main indicators of microbial oil
实施例4Example 4
以高山被包霉作为发酵菌种，在发酵所得发酵液中的含油率为5.50％时，加入发酵液质量0.03％的木犀草素，发酵完成后分别在70/80/90℃条件下对发酵液采用巴杀处理，所得发酵液分别经板框过滤得到浓缩菌体，然后将浓缩菌体烘干，加入己烷并升温至80℃，剪切搅拌，筛网过滤脱溶后得到二十碳四烯酸(ARA)毛油。该毛油经酸法脱胶、碱炼、脱色、脱臭后产出微生物油脂。所得产品中的主要成分见表4。When the oil content in the fermentation broth obtained from the fermentation is 5.50%, the luteolin with a fermentation broth quality of 0.03% is added, and the fermentation is carried out at 70/80/90 ° C respectively after the fermentation. The liquid is treated with bark, and the obtained fermentation broth is separately filtered through a plate frame to obtain concentrated cells, and then the concentrated cells are dried, hexane is added and heated to 80 ° C, sheared and stirred, and sieved to obtain 20 carbon. Tetraenoic acid (ARA) hair oil. The oil is degummed by acid method, alkali refining, decolorizing, and deodorizing to produce microbial oil. The main components in the obtained product are shown in Table 4.
表4 微生物油脂主要指标对比Table 4 Comparison of main indicators of microbial oil
实施例5Example 5
以高山被包霉作为发酵菌种，在发酵所得发酵液中的含油率为5.30％时，加入发酵液质量0.025％的木犀草素，发酵完成后在80℃条件下对发酵液采用巴杀处理，发酵液经板框过滤得到浓缩菌体，然后将浓缩菌体烘干，加入己烷并升温至80℃，剪切搅拌，筛网过滤脱溶后得到二十碳四烯酸(ARA)毛油。该毛油经酸法脱胶、碱炼、脱色、脱臭后产出微生物油脂。该微生物油脂再进一步在0.2Pa、185℃条件下，经分子蒸馏处理得到精制微生物油脂。所得产品中的主要成分见表5。When the oil content in the fermentation broth obtained from the fermentation is 5.30%, the luteolin with a mass of 0.025% of the fermentation broth is added, and after the fermentation is completed, the fermentation broth is treated at 80 ° C. The fermentation broth is filtered through a plate frame to obtain a concentrated bacterial cell, and then the concentrated bacterial cells are dried, hexane is added and the temperature is raised to 80 ° C, sheared and stirred, and the mesh is filtered and desolvated to obtain arachidonic acid (ARA) hair. oil. The oil is degummed by acid method, alkali refining, decolorizing, and deodorizing to produce microbial oil. The microbial oil and fat is further subjected to molecular distillation treatment at 0.2 Pa and 185 ° C to obtain a purified microbial fat. The main components in the obtained product are shown in Table 5.
表5 微生物油脂主要指标对比Table 5 Comparison of main indicators of microbial oil
实施例6Example 6
以高山被包霉作为发酵菌种，在发酵所得发酵液中的含油率为6.10％时，加入发酵液质量0.04％的木犀草素，发酵液经板框过滤得到浓缩菌体，浓缩菌体烘干后，加入己烷并升温至80℃，剪切搅拌，筛网过滤脱溶得到二十碳四烯酸(ARA)毛油。该毛油经酸法脱胶、碱炼、脱色、脱臭 后产出微生物油脂。所得微生物油脂中的主要成分见表6。When the oil content in the fermentation broth obtained by fermentation is 6.10%, the lardin is added with 0.04% of the fermentation broth, and the fermentation broth is filtered through the plate frame to obtain the concentrated bacteria, and the concentrated bacteria are dried. After drying, hexane was added and the temperature was raised to 80 ° C, sheared and stirred, and screen-filtered and desolvated to obtain arachidonic acid (ARA) hair oil. The oil is degummable by acid method, alkali refining, decolorization, and deodorization to produce microbial oil. The main components in the obtained microbial oil and fat are shown in Table 6.
表6 微生物油脂主要指标对比Table 6 Comparison of main indicators of microbial oil
实施例7Example 7
以裂殖壶藻作为发酵菌种，在发酵所得发酵液中的含油率为4.37％时，加入发酵液质量0.01％的木犀草素，发酵液经离心机浓缩脱水得到浓缩菌体，调节pH为8并加入碱性蛋白酶破壁处理，破壁后升温至80℃，离心分离得到二十二碳六烯酸(DHA)毛油，该毛油经酸法脱胶、碱炼、脱色、脱臭后产出微生物油脂。所得微生物油脂中的主要成分见表7。Using Schizophyllum algae as the fermentation strain, when the oil content in the fermentation broth obtained from the fermentation is 4.37%, luteolin is added in a 0.01% by mass of the fermentation broth, and the fermentation broth is concentrated and dehydrated by a centrifuge to obtain a concentrated bacterium, and the pH is adjusted to 8 and adding alkaline protease to break the wall treatment, after wall breaking, the temperature is raised to 80 ° C, and centrifuged to obtain docosahexaenoic acid (DHA) hair oil. The oil is degummed by acid method, alkali refining, decolorization and deodorization. Microbial oils are produced. The main components in the obtained microbial oil and fat are shown in Table 7.
表7 微生物油脂主要指标对比Table 7 Comparison of main indicators of microbial oil
实施例8Example 8
以双鞭甲藻作为发酵菌种，在发酵所得发酵液中的含油率为5.39％时，加入发酵液质量0.04％的齐墩果酸，发酵液经离心机浓缩脱水得到浓缩菌体，调节pH为8并加入碱性蛋白酶破壁处理，破壁后升温至80℃，离心分离得到二十二碳六烯酸(DHA)毛油，该毛油经酸法脱胶、碱炼、脱色、脱臭后产出微生物油脂。所得微生物油脂中的主要成分见表8。When dinoflagellate is used as the fermentation strain, when the oil content in the fermentation broth obtained by fermentation is 5.39%, the oleanolic acid of 0.04% of the fermentation liquid is added, and the fermentation liquid is concentrated and dehydrated by a centrifuge to obtain a concentrated bacteria to adjust the pH. 8 and adding alkaline protease to break the wall treatment, after wall breaking, the temperature was raised to 80 ° C, and centrifuged to obtain docosahexaenoic acid (DHA) hair oil. The oil was degummed by acid method, alkali refining, decolorization and deodorization. Produces microbial oils. The main components in the obtained microbial oil and fat are shown in Table 8.
表8 微生物油脂主要指标对比Table 8 Comparison of main indicators of microbial oil
实施例9Example 9
以双鞭甲藻作为发酵菌种，在发酵所得发酵液中的含油率为5.60％时，加入发酵液质量0.02％的木犀草素，发酵液经离心机浓缩脱水得到浓缩菌体，调节pH为8并加入碱性蛋白酶破壁处理，破壁后升温至80℃，离心分离得到二十二碳六烯酸(DHA)毛油，该毛油经酸法脱胶、碱炼、脱色、脱臭后产出微生物油脂。所得微生物油脂中的主要成分见表9。When dinoflagellate is used as a fermentation strain, when the oil content in the fermentation broth obtained by fermentation is 5.60%, luteolin is added to the fermentation liquid at a mass of 0.02%, and the fermentation liquid is concentrated and dehydrated by a centrifuge to obtain a concentrated bacteria, and the pH is adjusted to 8 and adding alkaline protease to break the wall treatment, after wall breaking, the temperature is raised to 80 ° C, and centrifuged to obtain docosahexaenoic acid (DHA) hair oil. The oil is degummed by acid method, alkali refining, decolorization and deodorization. Microbial oils are produced. The main components in the obtained microbial oil and fat are shown in Table 9.
表9 微生物油脂主要指标对比Table 9 Comparison of main indicators of microbial oil
实施例10Example 10
以微球藻作为发酵菌种，在破壁处理开始前加入发酵液质量0.02％的木犀草素，发酵液经板框过滤得到浓缩菌体，调节pH为8并加入碱性蛋白酶破壁处理。然后升温至80℃，离心分离处理后，提取得到二十二碳五烯酸(EPA)毛油。该毛油经酸法脱胶、碱炼、脱色、脱臭后产出微生物油脂。所得微生物油脂中的主要成分见表10。Microsphere algae was used as the fermentation strain, and luteolin was added to the fermentation liquid at a mass of 0.02% before the breaking treatment. The fermentation liquid was filtered through a plate to obtain a concentrated bacterial cell, and the pH was adjusted to 8 and the alkaline protease was broken. Then, the temperature was raised to 80 ° C, and after centrifugation, docosapentaenoic acid (EPA) hair oil was extracted. The oil is degummed by acid method, alkali refining, decolorizing, and deodorizing to produce microbial oil. The main components in the obtained microbial oil and fat are shown in Table 10.
表10 微生物油脂主要指标对比Table 10 Comparison of main indicators of microbial oils
实施例11Example 11
以微球藻作为发酵菌种，在破壁处理开始后0.5h时，加入发酵液质量0.03％的木犀草素，发酵液经板框过滤得到浓缩菌体，调节pH为11并加入碱性蛋白酶破壁处理。然后，然后升温至80℃，离心分离处理后提取得到二十二碳五烯酸(EPA)毛油。该毛油经酸法脱胶、碱炼、脱色、脱臭后产出微生物油脂。所得微生物油脂中的主要成分见表11。Microspheres were used as fermentation strains. At 0.5 h after the start of the wall breaking treatment, luteolin with a fermentation broth mass of 0.03% was added. The fermentation broth was filtered through a plate to obtain concentrated cells, and the pH was adjusted to 11 and alkaline protease was added. Broken wall treatment. Then, the temperature was raised to 80 ° C, and after extraction by centrifugation, docosapentaenoic acid (EPA) hair oil was extracted. The oil is degummed by acid method, alkali refining, decolorizing, and deodorizing to produce microbial oil. The main components in the obtained microbial oil and fat are shown in Table 11.
表11 微生物油脂主要指标对比Table 11 Comparison of main indicators of microbial oil
对比例1Comparative example 1
以与实施例1中同样的方法制备微生物油脂，其区别仅在于：不添加脂肪酶抑制剂，不进行分子蒸馏处理。该微生物油脂中的主要成分含量见表1。Microbial oil was prepared in the same manner as in Example 1 except that the lipase inhibitor was not added and no molecular distillation treatment was carried out. The main components in the microbial oil and fat are shown in Table 1.
对比例2Comparative example 2
以与实施例2中同样的方法制备微生物油脂，其区别仅在于：不添加脂肪酶抑制剂，不进行分子蒸馏处理。该微生物油脂中的主要成分含量见表2。A microbial fat was prepared in the same manner as in Example 2 except that the lipase inhibitor was not added and no molecular distillation treatment was carried out. The main components in the microbial oil and fat are shown in Table 2.
对比例3Comparative example 3
以与实施例3中同样的方法制备微生物油脂，其区别仅在于：不添加脂肪酶抑制剂，不进行分子蒸馏处理。该微生物油脂中的主要成分含量见表3。Microbial oil was prepared in the same manner as in Example 3 except that the lipase inhibitor was not added and no molecular distillation treatment was carried out. The content of the main components in the microbial oil is shown in Table 3.
对比例4Comparative example 4
以与实施例4中同样的方法制备微生物油脂，其区别仅在于：不添加脂肪酶抑制剂，不进行巴杀处理，该微生物油脂中的主要成分含量见表4。The microbial oil was prepared in the same manner as in Example 4 except that the lipase inhibitor was not added and the pasteurization treatment was not carried out. The contents of the main components in the microbial oil and fat are shown in Table 4.
对比例5Comparative example 5
以与实施例5中同样的方法制备微生物油脂，其区别仅在于：不添加脂肪酶抑制剂，不进行巴杀处理，不进行分子蒸馏处理，该微生物油脂中的主要成分含量见表5。Microbial fats and oils were prepared in the same manner as in Example 5 except that the lipase inhibitor was not added, the batter treatment was not carried out, and the molecular distillation treatment was not carried out. The contents of the main components in the microbial oil and fat are shown in Table 5.
对比例6Comparative example 6
以与实施例6中同样的方法制备微生物油脂，其区别仅在于：不添加脂肪酶抑制剂。该微生物油脂中的主要成分含量见表6。Microbial oil was prepared in the same manner as in Example 6, except that no lipase inhibitor was added. The main components in the microbial oil and fat are shown in Table 6.
对比例7Comparative example 7
以与实施例7中同样的方法制备微生物油脂，其区别仅在于：不添加脂肪酶抑制剂。该微生物油脂中的主要成分含量见表7。Microbial oil was prepared in the same manner as in Example 7 except that no lipase inhibitor was added. The main components in the microbial oil and fat are shown in Table 7.
对比例8Comparative example 8
以与实施例8中同样的方法制备微生物油脂，其区别仅在于：不添加脂肪酶抑制剂。该微生物油脂中的主要成分含量见表8。Microbial oil was prepared in the same manner as in Example 8 except that no lipase inhibitor was added. The main components in the microbial oil and fat are shown in Table 8.
对比例9Comparative example 9
以与实施例9中同样的方法制备微生物油脂，其区别仅在于：不添加脂肪酶抑制剂。该微生物油脂中的主要成分含量见表9。Microbial oil was prepared in the same manner as in Example 9 except that no lipase inhibitor was added. The main components in the microbial oil and fat are shown in Table 9.
对比例10Comparative example 10
以与实施例10中同样的方法制备微生物油脂，其区别仅在于：不添加脂肪酶抑制剂。该微生物油脂中的主要成分含量见表10。Microbial oil was prepared in the same manner as in Example 10 except that no lipase inhibitor was added. The contents of the main components in the microbial oil and fat are shown in Table 10.
对比例11Comparative Example 11
以与实施例11中同样的方法制备微生物油脂，其区别仅在于：不添加脂肪酶抑制剂。该微生物油脂中的主要成分含量见表11。Microbial oil was prepared in the same manner as in Example 11 except that no lipase inhibitor was added. The main components in the microbial oil and fat are shown in Table 11.
对比例12Comparative example 12
以与实施例4中同样的方法制备微生物油脂，其区别仅在于：不进行巴杀处理。该微生物油脂中的主要成分含量见表4。Microbial oil was prepared in the same manner as in Example 4 except that the pasteurization treatment was not carried out. The main components in the microbial oil and fat are shown in Table 4.
对比例13Comparative example 13
以与实施例5中同样的方法制备微生物油脂，其区别仅在于：不进行巴杀处理，不进行分子蒸馏处理。该微生物油脂中的主要成分含量见表5。Microbial fats and oils were prepared in the same manner as in Example 5 except that they were not subjected to a pulverization treatment and were not subjected to molecular distillation treatment. The content of the main components in the microbial oil is shown in Table 5.
对比例14Comparative example 14
以与实施例5中同样的方法制备微生物油脂，其区别仅在于：不进行巴杀处理。该微生物油脂中的主要成分含量见表5。Microbial oil was prepared in the same manner as in Example 5 except that the pasteurization treatment was not carried out. The content of the main components in the microbial oil is shown in Table 5.
对比例15Comparative example 15
以与实施例5中同样的方法制备微生物油脂，其区别仅在于：不进行分子蒸馏处理。该微生物油脂中的主要成分含量见表5。A microbial fat was prepared in the same manner as in Example 5 except that the molecular distillation treatment was not carried out. The content of the main components in the microbial oil is shown in Table 5.
由表1-9可知，采用相同的工艺处理方法，在发酵过程中添加脂肪酶 抑制剂后，能够明显的降低所得微生物油脂中的氯丙醇含量，但不会对微生物油脂中的目标油脂含量产生不良影响。并且，经巴杀处理和/或分子蒸馏处理，能够进一步降低微生物油脂中氯丙醇的含量。增加TAG的含量。It can be seen from Table 1-9 that the same process treatment method can significantly reduce the chloropropanol content in the obtained microbial oil after adding the lipase inhibitor in the fermentation process, but it does not target the target oil content in the microbial oil. Have an adverse effect. Further, the content of chloropropanol in the microbial oil can be further reduced by the killing treatment and/or the molecular distillation treatment. Increase the TAG content.
由表10和11可见，当在微生物菌种发酵结束后加入脂肪酶抑制剂时，虽然能够有效的降低微生物油脂中氯丙醇的含量，但同时也会降低微生物油脂中目标油脂的含量。It can be seen from Tables 10 and 11 that when a lipase inhibitor is added after the fermentation of the microbial strain, although the content of chloropropanol in the microbial oil can be effectively reduced, the content of the target fat in the microbial oil is also lowered.
由表1-3，5可见，分子蒸馏能够进一步降低油脂中的氯丙醇含量，并对油脂TAG含量有一定的提升作用。It can be seen from Tables 1-3 and 5 that molecular distillation can further reduce the chloropropanol content in the oil and fat, and has a certain effect on the TAG content of the oil.
由表4-5可见，巴杀处理能够进一步降低油脂中的氯丙醇含量，且可以明显提升油脂的TAG含量。As can be seen from Table 4-5, the pasteurization treatment can further reduce the chloropropanol content in the oil and fat, and can significantly increase the TAG content of the oil.
最后，本发明的方法仅为较佳的实施方案，并非用于限定本发明的保护范围。凡在本发明的精神和原则之内，所作的任何修改、等同替换、改进等，均应包含在本发明的保护范围之内。Finally, the method of the present invention is only a preferred embodiment and is not intended to limit the scope of the present invention. Any modifications, equivalent substitutions, improvements, etc. made within the spirit and scope of the present invention are intended to be included within the scope of the present invention.
本发明提供一种低氯丙醇微生物油脂及其制备方法。本发明通过在微生物菌种发酵的发酵液中添加脂肪酶抑制剂，经提取、精炼和脱臭处理，以降低所得微生物油脂中的缩水甘油酯和/或氯丙醇含量；其中，所述脂肪酶抑制剂能够抑制发酵液中微生物细胞内脂肪酶的活性。相比于现有技术中在降低氯丙醇含量时，需额外增设对应的处理过程，且最终产物中氯丙醇含量依然较高的技术问题，本发明的低氯丙醇微生物油脂及其制备方法，所得微生物油脂中氯丙醇含量低、处理工艺简单，且无需改变已有的工艺流程，具有较好的经济价值和应用前景。The invention provides a low chloropropanol microbial oil and a preparation method thereof. The invention reduces the glycidyl ester and/or chloropropanol content in the obtained microbial oil by adding a lipase inhibitor to the fermentation broth fermented by the microbial strain, and extracting, refining and deodorizing; wherein the lipase The inhibitor is capable of inhibiting the activity of lipase in the microbial cells in the fermentation broth. Compared with the prior art, in the case of reducing the chloropropanol content, it is necessary to additionally add a corresponding treatment process, and the technical problem of the chloropropanol content in the final product is still high, the low chloropropanol microbial oil of the invention and preparation thereof The method has the advantages that the obtained microbial oil has low chloropropanol content, simple treatment process, and no need to change the existing process flow, and has good economic value and application prospect.
Claims (10)
- 一种低氯丙醇微生物油脂，其特征在于，所述微生物油脂中的氯丙醇含量小于0.1ppm、TAG含量≥95％、DHA、ARA或EPA含量≥40％。A low chloropropanol microbial oil, characterized in that the microbial oil has a chloropropanol content of less than 0.1 ppm, an TAG content of ≥95%, and a DHA, ARA or EPA content of ≥40%.
- 一种低氯丙醇微生物油脂的制备方法，其特征在于：在微生物菌种发酵的发酵液中添加脂肪酶抑制剂；其中，所述脂肪酶抑制剂能够抑制发酵液中微生物细胞内脂肪酶的活性。A method for preparing a low-chlorinated propanol microbial oil, characterized in that a lipase inhibitor is added to a fermentation broth fermented by a microbial strain; wherein the lipase inhibitor can inhibit lipase in a microbial cell in a fermentation broth active.
- 如权利要求2所述的一种低氯丙醇微生物油脂的制备方法，其特征在于，所述脂肪酶抑制剂为：木犀草素、橙皮苷、新橙皮苷、枸橘苷、齐墩果酸中的一种或几种；优选为木犀草素。The method for preparing a microchloropropanol microbial oil according to claim 2, wherein the lipase inhibitor is: luteolin, hesperidin, neohesperidin, quercetin, and Qi Dun One or more of fruit acids; preferably luteolin.
- 如权利要求2或3所述的一种低氯丙醇微生物油脂的制备方法，其特征在于，在所述微生物菌种发酵的过程中，所述发酵液中的含油率达到4.0％-7.0％时，添加所述脂肪酶抑制剂；优选所述含油率达到5.0％-6.0％时，添加所述脂肪酶抑制剂。The method for preparing a microchloropropanol microbial oil according to claim 2 or 3, wherein in the fermentation of the microbial strain, the oil content in the fermentation liquid reaches 4.0% to 7.0%. The lipase inhibitor is added; preferably, the lipase inhibitor is added when the oil content is from 5.0% to 6.0%.
- 如权利要求2或3所述的一种低氯丙醇微生物油脂的制备方法，其特征在于，在所述微生物菌种发酵完成后、对所述微生物细胞进行破壁处理的0-1h内添加所述脂肪酶抑制剂，优选为破壁处理开始时立即添加。The method for preparing a microchloropropanol microbial oil according to claim 2 or 3, wherein after the fermentation of the microbial strain is completed, the microbial cell is subjected to a 0-1h breakage treatment. The lipase inhibitor is preferably added immediately after the start of the disruption treatment.
- 如权利要求2-5任一项所述的一种低氯丙醇微生物油脂的制备方法，其特征在于，所述脂肪酶抑制剂的添加量与所述发酵液的质量比为0.005％-0.05％，优选为0.02％-0.03％。The method for preparing a microchloropropanol microbial oil according to any one of claims 2 to 5, wherein the ratio of the amount of the lipase inhibitor added to the fermentation liquid is 0.005%-0.05. %, preferably from 0.02% to 0.03%.
- 如权利要求2-6任一项所述的一种低氯丙醇微生物油脂的制备方法，其特征在于，在所述微生物菌种发酵完成后，对发酵液进行巴杀处理；所述巴杀处理的温度为70-90℃，优选为80℃。The method for preparing a low-chlorinated propanol microbial oil according to any one of claims 2 to 6, wherein after the fermentation of the microbial strain is completed, the fermentation liquid is subjected to a pasteurization treatment; The temperature to be treated is 70-90 ° C, preferably 80 ° C.
- 如权利要求2-7任一项所述的一种低氯丙醇微生物油脂的制备方法，其特征在于，所述微生物菌种为：酵母、裂殖壶藻、双鞭甲藻、微球藻、破囊壶菌或高山被孢霉。The method for preparing a microchloropropanol microbial oil according to any one of claims 2 to 7, wherein the microbial strain is: yeast, Schizophyllum, Dinoflagellate, Microspheres , Thraustochytrium or Mortierella alpina.
- 如权利要求2-8任一项所述的一种低氯丙醇微生物油脂的制备方法，其特征在于，所述发酵液经提炼、精炼、脱臭处理后，再经分子蒸馏 处理，所述分子蒸馏处理的真空度为0-0.5Pa，优选为0.1-0.2Pa。The method for preparing a microchloropropanol microbial oil according to any one of claims 2-8, wherein the fermentation liquid is subjected to refining, refining, deodorization treatment, and then subjected to molecular distillation treatment, the molecule The degree of vacuum of the distillation treatment is from 0 to 0.5 Pa, preferably from 0.1 to 0.2 Pa.
- 如权利要求9所述的一种低氯丙醇微生物油脂的制备方法，其特征在于，所述分子蒸馏处理的温度为150-200℃，优选为180-190℃。A method for producing a low chloropropanol microbial fat according to claim 9, wherein the temperature of the molecular distillation treatment is from 150 to 200 ° C, preferably from 180 to 190 ° C.
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