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定制壳聚糖整合到硅藻土颗粒中作为固定化果糖基转移酶生产低聚果糖的支架。

Tailored chitosan integration in diatomaceous earth particles as a scaffold for fructosyltransferase immobilization in fructo-oligosaccharide production.

作者信息

Zhao Zishen, Xiao Ziqun, Jiang Bo, Chen Jingjing

机构信息

School of Food Science and Technology, Jiangnan University, Wuxi, China.

International Joint Laboratory on Food Safety, Jiangnan University, Wuxi, China.

出版信息

J Sci Food Agric. 2024 Aug 30;104(11):6563-6572. doi: 10.1002/jsfa.13480. Epub 2024 Apr 5.

Abstract

BACKGROUND

Fructo-oligosaccharide (FOS) belongs to the group of short inulin-type fructans and is one of the most important non-digestible bifid-oligosaccharides capable of biotransforming sucrose using fructosyltransferase (FTase). However, there are no immobilized FTase products that can be successfully used industrially. In this study, diatomite was subjected to extrusion, sintering and granulation to form diatomaceous earth particles that were further modified via chitosan aminomethylation for modification. FTase derived from Aspergillus oryzae was successfully immobilized on the modified support via covalent binding.

RESULTS

The immobilized enzyme activity was 503 IU g at an enzyme concentration of 0.6 mg mL, immobilization pH of 7.0 and contact time of 3 h. Additionally, the immobilization yield was 56.91%. Notably, the immobilized enzyme was more stable under acidic conditions. Moreover, the half-life of the immobilized enzyme was 20.80 and 10.96 times as long as that of the free enzyme at 45 and 60 °C, respectively. The results show good reusability, as evidenced by the 84.77% retention of original enzyme activity after eight cycles. Additionally, the column transit time of the substrate was 35.56 min when the immobilized enzyme was applied in a packed-bed reactor. Furthermore, a consistently high FOS production yield of 60.68% was achieved and maintained over the 15-day monitoring period.

CONCLUSIONS

Our results suggest that immobilized FTase is a viable candidate for continuous FOS production on an industrial scale. © 2024 Society of Chemical Industry.

摘要

背景

低聚果糖(FOS)属于短链菊粉型果聚糖,是最重要的非消化性双歧低聚糖之一,能够利用果糖基转移酶(FTase)对蔗糖进行生物转化。然而,目前尚无能够成功应用于工业生产的固定化FTase产品。在本研究中,对硅藻土进行挤压、烧结和造粒,形成硅藻土颗粒,再通过壳聚糖氨甲基化进行进一步改性。源自米曲霉的FTase通过共价结合成功固定在改性载体上。

结果

在酶浓度为0.6mg/mL、固定化pH为7.0、接触时间为3h的条件下,固定化酶活性为503IU/g。此外,固定化产率为56.91%。值得注意的是,固定化酶在酸性条件下更稳定。而且,在45℃和60℃时,固定化酶的半衰期分别是游离酶的20.80倍和10.96倍。结果显示出良好的可重复使用性,八个循环后仍保留84.77%的原始酶活性。此外,当将固定化酶应用于填充床反应器时,底物的柱通过时间为35.56分钟。此外,在为期15天的监测期内,FOS的生产产率始终保持在60.68%的高水平。

结论

我们的结果表明,固定化FTase是工业规模连续生产FOS的可行候选物。©2024化学工业协会。

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