Improving Enzyme-Assisted Extraction of Brazilin from Sappanwood (Caesalpinia Sappan L.) Extract by Fungal Cellulase

Abstract

Brazilin was one of phytoconstituent from sappanwood that widely used as textile-colouring agent and found in traditional concoction for therapeutic purposes. Due to obtain its maximum level in sappanwood (Caesalpinia sappan L.) extract, the extraction method of brazilin has been developed. Enzyme-assisted extraction (EAE) is one of green chemistry methods to achieve that outcome. This study aims to enhance brazilin level by optimize the EAE condition with fungi cellulase. The cellulase are produced by monoculture Aspergillus niger UICC371 in carboxymethyl cellulose submerged fermentation’s media. Sappanwood extracted with fungi cellulase through variation conditions: enzyme concentrations (2.0; 4,0; 6.0%); temperature (45, 50, 55℃); and time (1, 2, 3 hrs). The optimization are provided by response surface method-BoxBehnken design and brazilin level was carried out through High Performance Liquid Chromatography (HPLC) with asetonitril : 0,3% acetic acid in water (14.5 : 85.5) as eluents. The study showed that cellulase from monoculture of Aspergillus niger UICC371 showed optimum condition of cellulase-EAE method at 6.0% concentration enzyme at 50℃ for 3 hours extraction time which provide an increase in brazilin level to 5.014% compare to reflux method. Key Words: Caesalpinia sappan L., Cellulase, Enzyme assisted extraction, Fungi, Response surface method