In: Sydowia 76, (2024): 161-177; ISSN 0082-0598, DOI 10.12905/0380.sydowia76-2024-0161, Published online on July 16, 2024

Potential use of orange peel substrate to produce intracellular L-asparaginase by the endophyte Diaporthe sp. URM 8378

Ana Patrícia Sousa Lopes de Pádua, Leticia Francisca da Silva, Gualberto Segundo Agamez-Montalvo, Jadson Diogo Pereira Bezerra, Adalberto Pessoa Júnior, Cristina Maria de Souza-Motta & Keila Aparecida Moreira*

1 Departamento de Micologia Prof. Chaves Batista, Universidade Federal de Pernambuco (UFPE). Rua Prof. Nelson Chaves, s/n – Cidade Universitária, CEP: 50670-901. Recife, Pernambuco, Brazil
2 Departamento de Estatística e Matemática Aplicada, Universidade Federal do Ceará (UFC). Av. Mister Hull, s/n, Pici, CEP: 60455-760. Fortaleza, Ceará, Brazil
3 Laboratório de Micologia, Departamento de Biociências e Tecnologia, Instituto de Patologia Tropical e Saúde Pública (IPTSP), Universidade Federal de Goiás (UFG). Rua 235, s/n, Setor Universitário, CEP: 74605-050. Goiânia, Goiás, Brazil
4 Departamento de Bioquímica e Tecnologia Farmacêutica, Universidade de São Paulo. Av. Prof. Lineu Prestes, 580 – Bloco 16. CEP: 05508-000. São Paulo, São Paulo, Brazil
5 Laboratório de Microbiologia, Tecnologia Enzimática e Bioprodutos, Universidade Federal do Agreste de Pernambuco (UFAPE). Av. Bom Pastor, s/n. Boa Vista, CEP: 55292-278. Garanhuns, Pernambuco, Brazil

Pádua A.P.S.L., Silva L.F., Agamez-Montalvo G.S., Bezerra J.D.P., Pessoa Júnior A., Souza-Motta C.M. & Moreira K.A. (2023) Potential use of orange peel substrate to produce intracellular L-asparaginase by the endophyte Diaporthe sp. URM 8378. – Sydowia 76: 161–177.

This study aimed to select an agro-industrial substrate with low cost and efficiency for the production of L-asparaginase (LASNase) by the endophyte Diaporthe sp. URM 8378 using submerged fermentation and optimized enzymatic production. Fungal growth and L-asparaginase production curves were generated. A factorial experimental design (23) was carried out, and enzyme production was analysed using pH, substrate (%), and inoculum concentration (g) as variables. Orange peel flour was found to be the best inducer, and Diaporthe sp. URM 8378 produced a maximum intracellular yield for L-asparaginase of 2.56 U/g, followed by 0.85 U/g using eggshell flour, 0.80 U/g with L-asparagine (control) and 0.74 U/g with L-proline. The highest biomass production by the fungus (0.72 g/ml) occurred after 144 hours of cultivation at pH 2.5, and the best L-asparaginase production (1.05 U/g) occurred after 48 hours of fermentation at pH 4.8. The production of L-asparaginase was partially optimized between 2.56 U/g to 3.46 U/g at pH 6.0, inoculum of 1.5 g and orange peel flour concentration of 1.5 %, an increase of 26.02 % in relation to the initial production. The orange peel flour proved to be effective in increasing the L-sparaginase production of the endophyte Diaporthe sp. URM 8378.

Keywords: Agro-industrial waste, anticancer enzyme, Diaporthe sp., highest enzyme productivity, submerged fermentation.