三相分離法食用菇廢棄固態培養基萃取物之漆酵素對合成染料降解之研究

Degradation of synthetic dyes by laccase enzymes extracting from edible mushroom waste solid culture by three-phase separation method

詹廷寬(Quentin Zhan)

指導教授 ： 周崇榮

中原大學/工學院/化學工程研究所/碩士(2019年)

https://doi.org/10.6840/cycu201900678

摘要

在現代這個追求環境保護的綠色工業時代，酵素工程為近年來注重的環保綠色技術，因此本研究注重於將台灣食用菇種殖農業常見的廢棄太空包(食用菇固態廢棄培養基)，經由簡單的粗萃及三相分離程序純化後，取得濃度較高之酵素，並探討酵素運用在染料處理上。
本研究經過蛋白質定量、活性測試及SDS PAGE定性廢棄太空包萃取液，萃取液對四種染料處理：Methyl Orange、Methyl Blue、Acid Orange 7、Acid Blue 25，其中Methyl Orange、Acid Orange 7為偶氮染料，Methyl Blue為芳基染料，Acid Blue 25為蒽醌染料，並且分別加入兩種氧化還原介體：ABTS、過氧化氫。以動力學曲線和高效液相層析儀檢測。
結果顯示，經過三相分離純化後，杏鮑菇(Pleurotus eryngii)之蛋白質濃度為 6.64 mg/mL，秀珍菇(Pleurotus ostreatus) 之蛋白質濃度為 9.67 mg/mL，萃取液內含漆酶和錳過氧化物酶。在動力學參數的部分，可以發現杏鮑菇萃取液的Kmax在四種染料都是偏低的，秀珍菇萃取液的Vm在四種染料都是偏高的，Methyl Orange、Acid Orange 7在被加入ABTS都能有穩定的降解，Methyl Blue受H2O2影響很大，三種酵素在處理Acid Blue 25都有很好的表現，在杏鮑菇萃取液處理Acid Blue 25染料上，未加入ABTS有近似於加入ABTS的效果，而在秀珍菇萃取液處理Acid Orange 7染料上，未加入ABTS比加入ABTS更好的效果，或許是萃取液其中內含了天然的氧化還原介體，促進了氧化還原能力。

關鍵字

漆酵素 ； 錳過氧化物酶 ； 氧化還原介體 ； 偶氮染料 ； 廢棄固態培養基 ； 三相分離

並列摘要

Enzyme process is an environmentally-friendly technology that has been focuses on in recent years. In textile industry, dye-containing wastewater has to be processed before discharged. Therefore, a simple and affordable source for enzymes that can degrade synthetic dyes would be important for green economy. In this study, we focuses on recycling of the solid state cultivation capsules commonly used in Taiwan's edible mushroom production. Enzyme mixture can be partially purified by a simple three-phase separation procedure (TPP) for dye treatment.
Subsequently, protein quantification, activity test and SDS PAGE were used to characterize the extracts of Pleurotus eryngii (P.e.) and Pleurotus ostreatus (P.o.). Moreover, two redox mediators were added separately: ABTS, hydrogen peroxide treatment to evaluate their impacts on process efficiency. The kinetic analysis and high performance liquid chromatography (HPLC) were used for quantitative assay. The extract and the commercially available laccase are used to treat four dyes: Methyl Orange, Acid Orange 7 (azo dyes), Methyl Blue (triarylmethyl dye), and Acid Blue 25 (anthraquinone dye).
The results showed that the protein concentration of extracts of P.e. and P.o. was 6.64 mg/mL and 9.67 mg/mL, respectively. In the kinetic parameters, it can be found that the Km of P.e. extract is low in all four dyes, and the Vm of P.o. extract is high in all four dyes, Methyl Orange and Acid Orange 7 could be acceleratedlly degraded by these extracts. Enzymes performed well to degrade Acid Blue 25 while Methyl blue was not affected significantly. Optimize strategy based on the experimental data was proposed for each dye with a proper combination of enzyme extracts and redox mediator.

並列關鍵字

laccase ； manganese peroxidase ； redox mediator ； azo dye ； waste solid medium ； three-phase separation

參考文獻

參考文獻

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