Optimization of medium parameters by response surface methodology (RSM) for enhanced production of cutinase from Aspergillus sp. RL2Ct.

Vijaya Kumari,Vijay Kumar,Tek Chand Bhalla,Ravin Chauhan,Mohammad Asif

doi:10.1007/s13205-016-0460-4

Abstract

Cutinases are hydrolytic enzymes which catalyzes esterification and trans-esterification reactions that make them highly potential industrial biocatalyst. In the present investigation microorganisms showing cutinase activity were isolated from plant samples. The strain showing maximum cutinase activity was identified by 18S rDNA sequencing as Aspergillus sp. RL2Ct and was selected for further studies. To achieve maximum enzyme production, the medium components affecting cutinase production were screened by Plackett–Burman followed by central composite design. The results obtained suggested that cutin, temperature and CaCl2 have influenced the cutinase production significantly with very high confidence levels. Cutinase production was maximum (663 U/mg protein) when using cutin prepared from orange peel as sole source of carbon. An overall 4.33-fold increase in the production of cutinase was observed after optimization of culture conditions (including 2.5-fold increase using RSM) during 24 h of incubation. The production time of Aspergillus sp. RL2Ct cutinase is significantly lower than the most of the earlier reported cutinase-producing fungus.

Highlights

The cuticle protects the leaves, fruits and other softer parts of plant from dehydration and it acts as the outermost barrier in plant parts to infestation by pathogens
The maximum cutinase activity was recorded in fungal isolate RL2Ct (153 ± 3.0 U/mg protein)
The 18S rDNA gene sequence of this fungal isolate was carried out and the obtained sequence was deposited in the GenBank database with accession no

Summary

Introduction

The cuticle protects the leaves, fruits and other softer parts of plant from dehydration and it acts as the outermost barrier in plant parts to infestation by pathogens. Cutinases have been used as important catalyst to produce dehydrated fruits, dairy products, flavoring compounds and some important fatty acids such as eicosapentanoic acid (EPA) and docosahexanoic acid (DHA) (Dutta et al 2009). These enzymes have been used to degrade various insoluble polymer films like poly(ethylene terephthalate) (PETP) and poly(e-caprolactone) (PCL) (Murphy et al 1996; Liu et al 2009; Korpecka et al 2010; Greimel et al 2013). Fungal cutinases are considered as being intermediate between lipases and esterases (Carvalho et al 1999; Longhi and Cambillau 1999)

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Discussion

Conclusion