Citrus limetta peels: a promising substrate for the production of multienzyme preparation from a yeast consortium

Abstract

Citrus limetta peels (CLP), a waste material generated by juice industries, has scarcely been reported for the production of yeast enzymes. The study was conducted to obtain a multienzyme preparation from a yeast consortium under solid-state fermentation of CLP. The substrate, CLP, was pretreated using either acid or alkali, and factors affecting production of multienzyme were studied by generating two separate Plackett–Burman designs. Since, alkali-pretreated CLP yielded higher titers; therefore, significant factors affecting multienzyme preparation using this substrate were optimized by employing Box–Behnken design. The analysis revealed that under optimized conditions, i.e., cultivation of yeast strains for 72 h to alkali-pretreated CLP moistened with mineral salt medium having pH 5 yielded more than 10 IU mL⁻¹ of cellulase, xylanase, and amylase. The multienzyme was studied for its application to saccharify fruit and non-fruit wastes and for orange juice clarification. The data showed that the enzyme preparation could release 3.03 mg L⁻¹ h⁻¹ of reducing sugars from various crude substrates and was able to reduce turbidity of orange juice by 11% with substantial decrease in viscosity and acidity. Hence, CLP appeared as a promising substrate to produce multienzyme preparation from yeast consortium.

Keywords

Box–Behnken design / Citrus limetta / Multienzyme preparation / Plackett–Burman design / Saccharification / Yeast consortium

References

[1] Ahmed A, Sohail M. Characterization of pectinase from Geotrichum candidum AA15 and its potential application in orange juice clarification. J King Saud Univ Sci, 2019

[2] Ahmed A, Khan MN, Ahmad A, . Optimization of pectinase production from Geotrichum candidum AA15 using response surface methodology. Pak J Bot, 2019, 51: 743-750.

[3] Arshad ZIM, Amid A, Yusof F, . Bromelain: an overview of industrial application and purification strategies. Appl Microbiol Biotechnol, 2014, 98: 7283-7297.

[4] Brijwani K, Oberoi HS, Vadlani PV. Production of a cellulolytic enzyme system in mixed-culture solid-state fermentation of soybean hulls supplemented with wheat bran. Process Biochem, 2010

[5] Chew KW, Yap JY, Show PL, . Microalgae biorefinery: high value products perspectives. Bioresour Technol, 2017

[6] Cui F, Zhao L. Optimization of xylanase production from Penicillium sp. WX-Z1 by a two-step statistical strategy: Plackett–Burman and Box–Behnken experimental des. Int J Mol Sci, 2012

[7] Deswal D, Khasa YP, Kuhad RC. Optimization of cellulase production by a brown rot fungus Fomitopsis sp. RCK2010 under solid state fermentation. Bioresour Technol, 2011

[8] Ferreira JA, Mahboubi A, Lennartsson PR, Taherzadeh MJ. Waste biorefineries using filamentous ascomycetes fungi: present status and future prospects. Bioresour Technol, 2016, 215: 334-345.

[9] Grohmann K, Cameron RG, Buslig BS. Fractionation and pretreatment of orange peel by dilute acid hydrolysis. Bioresour Technol, 1995

[10] Herrero AA, Gomez RF, Roberts MF. 31P NMR studies of Clostridium thermocellum. Mechanism of end product inhibition by ethanol. J Biol Chem, 1985, 260: 7442-7451.

[11] Hmad IB, Abdeljalil S, Saibi W, . Medium initial pH and carbon source stimulate differential alkaline cellulase time course production in Stachybotrys microspora. Appl Biochem Biotechnol, 2014

[12] Lee WC, Yusof S, Hamid NSA, Baharin BS. Optimizing conditions for enzymatic clarification of banana juice using response surface methodology (RSM). J Food Eng, 2006

[13] Li PJ, Xia JL, Shan Y, Nie ZY. Comparative study of multi-enzyme production from typical agro-industrial residues and ultrasound-assisted extraction of crude enzyme in fermentation with Aspergillus japonicus PJ01. Bioprocess Biosyst Eng, 2015

[14] Lynd LR, Weimer PJ, van Zyl WH, Pretorius IS. Microbial cellulose utilization: fundamentals and biotechnology. Microbiol Mol Biol Rev, 2002

[15] Madrera RR, Bedriñana RP, Valles BS. Production and characterization of aroma compounds from apple pomace by solid-state fermentation with selected yeasts. LWT Food Sci Technol, 2015

[16] Manan MA, Webb C. Modern microbial solid state fermentation technology for future biorefineries for the production of added-value products. Biofuel Res J, 2017, 4: 730-740.

[17] Mantzouridou FT, Paraskevopoulou A, Lalou S. Yeast flavour production by solid state fermentation of orange peel waste. Biochem Eng J, 2015

[18] Miller GL. Use of dinitrosalicylic acid reagent for determination of reducing sugar. Anal Chem, 1959

[19] Nagar S, Mittal A, Gupta VK. Enzymatic clarification of fruit juices (apple, pineapple, and tomato) using purified Bacillus pumilus SV-85S xylanase. Biotechnol Bioprocess Eng, 2012

[20] Naseeb S, Sohail M, Ahmad A, Khan SA. Production of xylanases and cellulases by Aspergillus fumigatus MS16 using crude lignocellulosic substrates. Pak J Bot, 2015, 47: 779-784.

[21] Oberoi HS, Vadlani PV, Madl RL, . Ethanol production from orange peels: two-stage hydrolysis and fermentation studies using optimized parameters through experimental design. J Agric Food Chem, 2010

[22] Oberoi HS, Vadlani PV, Nanjundaswamy A, . Enhanced ethanol production from Kinnow mandarin (Citrus reticulata) waste via a statistically optimized simultaneous saccharification and fermentation process. Bioresour Technol, 2011

[23] Pirzadah T, Garg S, Singh J, . Characterization of Actinomycetes and Trichoderma spp. for cellulase production utilizing crude substrates by response surface methodology. Springerplus, 2014

[24] Qadir F, Shariq M, Ahmed A, Sohail M. Evaluation of a yeast co-culture for cellulase and xylanase production under solid state fermentation of sugarcane bagasse using multivariate approach. Ind Crops Prod, 2018

[25] Rahnama N, Mamat S, Shah UKM, . Effect of alkali pretreatment of rice straw on cellulase and xylanase production by local Trichoderma harzianum SNRS3 under solid state fermentation. BioResources, 2013

[26] Raimbault M. General and microbiological aspects of solid substrate fermentation. Electron J Biotechnol, 1998, 1: 1-15.

[27] Rehman S, Aslam H, Ahmad A, . Production of plant cell wall degrading enzymes by monoculture and co-culture of Aspergillus niger and Aspergillus terreus under SSF of banana peels. Brazilian J Microbiol, 2014, 45: 1485-1492.

[28] Rosyida VT, Indrianingsih AW, Maryana R, Wahono SK. Effect of temperature and fermentation time of crude cellulase production by Trichoderma reesei on straw substrate. Energy Procedia, 2015, 65: 368-371.

[29] Sadhu S. Cellulase production by bacteria: a review. Br Microbiol Res J, 2013

[30] Sagar NA, Pareek S, Sharma S, . Fruit and vegetable waste: bioactive compounds, their extraction, and possible utilization. Compr Rev Food Sci Food Saf, 2018

[31] Salihu A, Alam MZ, Abdulkarim MI, Salleh HM. Optimization of lipase production by Candida cylindracea in palm oil mill effluent based medium using statistical experimental design. J Mol Catal B Enzym, 2011

[32] Sandhu SK, Oberoi HS, Dhaliwal SS, . Ethanol production from Kinnow mandarin (Citrus reticulata) peels via simultaneous saccharification and fermentation using crude enzyme produced by Aspergillus oryzae and the thermotolerant Pichia kudriavzevii strain. Ann Microbiol, 2012

[33] Sankaran R, Bong JH, Chow YH, . Reverse micellar system in protein recovery—a review of the latest developments. Curr Protein Pept Sci, 2019

[34] Shariq M, Sohail M. Application of Candida tropicalis MK-160 for the production of xylanase and ethanol. J King Saud Univ Sci, 2018

[35] Shariq M, Sohail M. Production of cellulase and xylanase from Candida tropicalis (MK-118) on purified and crude substrates. Pak J Bot, 2020

[36] Shariq M, Muhammad F, Ahmad A, . Production and characterization of endoglucanase from an indigenous yeast strain. Pak J Bot, 2018, 50: 2413-2421.

[37] Sharma PK. Optimization of process parameters for fruit juice clarification using silica immobilized xylanase from Pseudomonas sp. Pure Appl Biol, 2012

[38] Sharma S, Sharma J, Mandhan RP. Lucrative pectinase production by novel strain Pseudozyma sp. SPJ with statistical optimization techniques using agro-industrial residues. Front Biol, 2014

[39] Song HT, Gao Y, Yang YM, . Synergistic effect of cellulase and xylanase during hydrolysis of natural lignocellulosic substrates. Bioresour Technol, 2016

[40] Sutherland IW. Biofilm exopolysaccharides: a strong and sticky framework. Microbiology, 2001, 147: 3-9.

[41] Suwannarangsee S, Arnthong J, Eurwilaichitr L, Champreda V. Production and characterization of multi-polysaccharide degrading enzymes from Aspergillus aculeatus BCC199 for saccharification of agricultural residues. J Microbiol Biotechnol, 2014

[42] Tariq R, Ansari I, Qadir F, . Optimization of endoglucanase production from thermophilic strain of Bacillus licheniformis RT-17 and its application for saccharification of sugarcane bagasse. Pak J Bot, 2018, 50: 807-816.

[43] Yahya S, Jahangir S, Shaukat SS, . Production optimization by using Plackett-Burman design and partial characterization of amylase from Aspergillus tubingensis SY 1. Pak J Bot, 2016, 48: 2557.

[44] Yasin Ashraf M, Ashraf M, Akhtar M, . Improvement in yield, quality and reduction in fruit drop in Kinnow (Citrus reticulata Blanco) by exogenous application of plant growth regulators, potassium and zinc. Pak J Bot, 2013, 45: 433-440.

[45] Yoon LW, Ang TN, Ngoh GC, Chua ASM. Fungal solid-state fermentation and various methods of enhancement in cellulase production. Biomass Bioenergy, 2014, 67: 319-338.

[46] Zhang C, Xing XH, Liu MS. Production of multienzymes consisting of alkaline amylase and cellulase by mixed alkalophilic culture and their potential use in the saccharification of sweet potato. Biochem Eng J, 2004

Funding

Third World Academy of Sciences(10-134)