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固定化白腐真菌小球对偶氮染料的脱色作用。

Decolouration of azo dyes by Phanerochaete chrysosporium immobilised into alginate beads.

机构信息

Department of Chemical Engineering, Rovira i Virgili University, 43007, Tarragona, Spain.

出版信息

Environ Sci Pollut Res Int. 2010 Jan;17(1):145-53. doi: 10.1007/s11356-009-0109-5. Epub 2009 Mar 4.

DOI:10.1007/s11356-009-0109-5
PMID:19259719
Abstract

BACKGROUND, AIM AND SCOPE: Because of high discharged volumes and effluent composition, wastewater from the textile industry can be considered as the most polluting amongst all industrial sectors, thus greatly requiring appropriate treatment technologies. Although some abiotic methods for the reduction of several dyes exist, these require highly expensive catalysts and reagents. Biotechnological approaches were proven to be potentially effective in the treatment of this pollution source in an eco-efficient manner. The white-rot fungi are, so far, the most efficient microorganisms in degrading synthetic dyes. This white-rot fungi's property is due to the production of extracellular lignin-modifying enzymes, which are able to degrade a wide range of xenobiotic compounds because of their low substrate specificity. In this paper, we studied the ability of the white-rot fungus Phanerochaete chrysosporium immobilised into Ca-alginate beads to decolourise different recalcitrant azo dyes such as Direct Violet 51 (DV), Reactive Black 5 (RB), Ponceau Xylidine (PX) and Bismark Brown R (BB) in successive batch cultures. To the best of our knowledge, this is the first study on the immobilisation of P. chrysosporium into Ca-alginate beads for its application in dye decolouration.

MATERIALS AND METHODS

P. chrysosporium was immobilised into Ca-alginate beads using a method of gel recoating to minimise cellular leaking. The immobilised fungus was transferred to 250-ml Erlenmeyer flasks containing 50 ml of growth medium and incubated on an orbital shaker at 150 rpm and 30 degrees C for 7 days. The ratio of beads/medium used was 10% (w/v). The dyes were added into the culture flasks when MnP production started (50 U l(-1)), which corresponded with the seventh cultivation day. MnP activity and dye decolouration were measured spectrophotometrically.

RESULTS

The dyes DV, RB and PX were almost totally decolourised at the end of each batch during the course of three successive batches. However, the dye BB was more resistant to decolouration and it was not completely decolourised (86.7% in 144 h). Further, the beads were kept in sterilised calcium chloride (2 g l(-1)) for 3 weeks at 4 degrees C. After these three storage weeks, the immobilised P. chrysosporium was again efficiently reused for azo dye decolouration during two successive batches, decolouration being more effective even for BB. Also, the in vitro decolouration of the aforementioned azo dyes by crude MnP from P. chrysosporium was performed. The decolouration levels obtained were lower than those attained with the whole cultures especially for RB and BB dyes, in spite of the fact that dye concentrations used were considerable lower.

DISCUSSION

The good performance of the immobilisation system was likely due to the gel re-coating method utilised to prepare the alginate beads which not only maintained the beads integrity but also avoided cellular leaking. The lower decolouration percentages obtained by the enzyme indicates that the mycelial biomass may supply other intracellular or mycelial-bound enzymes, or other compounds that favour dye decolouration.

CONCLUSIONS

Immobilised P. chrysosporium efficiently decolourised different types of azo dyes. In this decolouration process, the MnP secreted by the fungus played the main role whilst adsorption was found to be negligible except for the dye BB.

RECOMMENDATIONS AND PERSPECTIVES

Efforts should be made to scale up and apply fungal decolouration techniques to real industrial dye-containing wastewater. Further, detailed characterisation of the intermediates and metabolites produced during biodegradation must be done to ensure the safety of the decolourised wastewater.

摘要

背景、目的和范围:由于排放量高和废水成分复杂,纺织工业废水被认为是所有工业部门中污染最严重的废水,因此需要采用适当的处理技术。尽管存在一些用于减少多种染料的非生物方法,但这些方法需要使用非常昂贵的催化剂和试剂。生物技术方法已被证明是一种在生态效率方面有效处理这种污染源的方法。白腐真菌迄今为止是降解合成染料最有效的微生物。白腐真菌之所以具有这种特性,是因为它能够产生能够降解多种外来化合物的胞外木质素修饰酶,这些酶的底物特异性较低。在本文中,我们研究了固定在 Ca-藻酸盐珠中的白腐真菌 Phanerochaete chrysosporium 对不同难降解偶氮染料(如直接紫 51(DV)、活性黑 5(RB)、对位红(PX)和俾斯麦棕 R(BB))的脱色能力,这些染料在连续批次培养中使用。据我们所知,这是首次研究将 P. chrysosporium 固定在 Ca-藻酸盐珠中用于染料脱色的应用。

材料和方法

使用凝胶再涂层的方法将 P. chrysosporium 固定在 Ca-藻酸盐珠中,以最小化细胞渗漏。将固定化真菌转移到装有 50 ml 生长培养基的 250 毫升 Erlenmeyer 摇瓶中,并在 30 摄氏度和 150 rpm 的条件下在摇床上培养 7 天。使用的珠/培养基比例为 10%(w/v)。当 MnP 生产开始(50 U l(-1))时,即在第七天的培养过程中,将染料添加到培养瓶中。通过分光光度法测量 MnP 活性和染料脱色。

结果

在三个连续批次的过程中,当 MnP 生产开始时(50 U l(-1)),即第七天的培养过程中,将染料添加到培养瓶中。在三个连续批次的过程中,当 MnP 生产开始时(50 U l(-1)),即第七天的培养过程中,将染料添加到培养瓶中。DV、RB 和 PX 等染料在每个批次结束时几乎完全脱色。然而,BB 染料更难脱色,没有完全脱色(144 小时内脱色 86.7%)。此外,将珠子在 4°C 的 2 g l(-1)的无菌氯化钙中保存 3 周。经过这三个储存周后,固定化的 P. chrysosporium 再次在两个连续批次中有效重复用于偶氮染料的脱色,甚至对 BB 染料的脱色效果更好。此外,还进行了来自 P. chrysosporium 的粗 MnP 对上述偶氮染料的体外脱色。与整个培养物相比,获得的脱色水平较低,特别是对于 RB 和 BB 染料,尽管使用的染料浓度要低得多。

讨论

固定化系统的良好性能可能归因于用于制备藻酸盐珠的凝胶再涂层方法,该方法不仅保持了珠子的完整性,而且避免了细胞渗漏。酶的脱色率较低表明,菌丝体生物量可能提供其他细胞内或菌丝体结合的酶,或其他有利于染料脱色的化合物。

结论

固定化的 P. chrysosporium 有效地对不同类型的偶氮染料进行脱色。在这个脱色过程中,真菌分泌的 MnP 起主要作用,而吸附作用可以忽略不计,除了 BB 染料。

建议和展望

应努力将真菌脱色技术放大并应用于实际的工业含染料废水处理中。此外,必须对生物降解过程中产生的中间产物和代谢物进行详细的表征,以确保脱色废水的安全性。

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