Elsevier

Food Chemistry

Volume 124, Issue 3, 1 February 2011, Pages 1096-1098
Food Chemistry

Short communication
Enzymatic hydrolysis of capsaicins for the production of vanillylamine using ECB deacylase from Actinoplanes utahensis

https://doi.org/10.1016/j.foodchem.2010.06.070Get rights and content

Abstract

Echinocandin acylase from Actinoplanes utahensis NRRL 12052 (ECB deacylase) was used for the hydrolysis of capsaicins (trans-8-methyl-N-vanillyl-6-nonenamide and 8-methyl-N-vanillyl-6-nonanamide) into vanillylamine. Caspaicins were extracted from Capsicum annuum var. Red Cayenne. The enzyme was partially purified and used for improving molar conversion and reaction rate of the biotransformation. Under optimised conditions, the preparative biotransformation of 8.2 mM capsaicins on 200 ml scale allowed for 92% molar conversions and 59% of recovered vanillylamine.

Introduction

Chily peppers and related plants of the Capsicum family produce a mixture of capsaicins (trans-8-methyl-N-vanillyl-6-nonenamide and 8-methyl-N-vanillyl-6-nonanamide), which are the most important molecules in terms of abundance and pungency (Suzuki & Iwai, 1984). Deacylation of these amides furnishes vanillylamine (4-hydroxy-3-methoxybenzylamine) which can be further enzymatically converted into vanillin using specific microbial or plant oxidase (Sudhakar Johnson et al., 1996, Yoshida et al., 1997). This two-step enzymatic conversion of natural capsaicins into vanillin (Scheme 1) is attractive since the product obtained can be classified as a natural flavour. Indeed, US and European legislations state that ‘natural’ flavour substances can only be prepared either by extraction from natural sources or by enzymatic or microbial processes, which involve precursors isolated from nature (Serra, Fuganti, & Brenna, 2005).

Candida antarctica lipase B (CALB) (Reyes Duarte et al., 2000, Torres-Gavilan et al., 2006) and with an extracellular enzyme from Streptomyces mobaraensis (Koreishi et al., 2006) with best yields around 2.3 mM vanillylamine after 72 h. In this work, we have screened twenty actinomycetes known as producers of acylase activity (Gandolfi, Jovetic, Marinelli, & Molinari, 2007) for selecting enzymes able to catalyse the hydrolysis of capsaicins with high molar conversions. An acylase produced from Actinoplanes utahensis NRRL 12052 (Boeck et al., 1988, Kreuzman A:J. et al., 2000) was selected and the enzymatic transformation was optimised.

Section snippets

Chemicals

Unless otherwise stated, all reagents were of analytical grade from Sigma (St. Louis, MO).

Microorganisms

The actinomycetes used in this study (Gandolfi et al., 2007) were from our collection and from commercial collections (NRRL, Northern Utilisation Research Branch, USDA, Peoria, Illinois, USA and ATCC-LGC Promochem, American Type Culture Collection, LGC Promochem, Sesto San Giovanni, Milano, Italy). Spores of actinomycetes were maintained in Nutrient Glycerol (Nutrient broth Difco 8 g l−1, glycerol 150 ml l−1)

Results

A screening for the hydrolysis of capsaicins was performed amongst 20 actinomycetes previously assayed for acylase activity (Gandolfi et al., 2007), showing that A. utahensis NRRL 12052 was the only strain able to furnish molar conversion above 15%. The biotransformations were carried out using the mixture of capsaicins (capsaicin/dihydrocapsaicin 65/35) extracted from C. annuum (Table 1).

The activity was mostly mycelium-bound and both the capsaicins were hydrolysed with similar reaction rates

Conclusions

In conclusion, we have described a novel method for the efficient bioconversion of capsaicins into vanillylamine using the membrane-bound ECB deacylase from A. utahensis. The enzyme shows better results in terms of rates and conversions than what observed with commercial CALB lipase (Torres-Gavilan et al., 2006) and an acylase from Streptomyces mobaraensis (Koreishi et al., 2006); moreover, the set-up of preparative biotransformation using ECB deacylase as partially purified enzymatic

Acknowledgements

This work has been supported by Ministry of University and Research (MIUR) funding PRIN: “Production and application of plant and microbial enzymes for the synthesis of vanillin”.

References (12)

There are more references available in the full text version of this article.

Cited by (0)

View full text