Optimization of glutaminase-free L-asparaginase production using mangrove endophytic Lysinibacillus fusiformis B27

Abstract

Background: The mangrove, Rhizophora mucronata, an essential source of endophytic bacteria, was investigated for its ability to produce glutaminase-free L-asparaginase. The study aimed to obtain glutaminase-free L-asparaginase-producing endophytic bacteria from the mangrove and to optimize enzyme production. Methods: The screening of L-asparaginase-producing bacteria used modified M9 medium. The potential producer was further analyzed with respect to its species using 16S rRNA gene sequencing. Taguchi experimental design was applied to optimize the enzyme production. Four factors (L-asparagine concentration, pH, temperature, and inoculum concentration) were selected at four levels. Results: The results indicated that the endophytic bacteria Lysinibacillus fusiformis B27 isolated from R. mucronata was a potential producer of glutaminase-free L-asparaginase. The experiment indicated that pH 6, temperature at 35°C, and inoculum concentration of 1.5% enabled the best production and were essential factors. L-asparagine (2%) was less critical for optimum production. Conclusions: L. fusiformis B27, isolated from Rhizophora mucronata, can be optimized for L-ASNase enzyme production using optimization factors (L-ASNase, pH, temperature, and inoculum), which can increase L-ASNase enzyme production by approximately three-fold.

Desirable L-asparaginases for treating cancer and current research trends

“… Ashok et al (2019) screened fungal species from Antarctic soil and moss that generate L-asparaginase devoid of glutaminase and urease ( Ashok et al, 2019 ). Glutaminase-free L-ASNase has been tested in numerous investigations from various sources ( El-naggar et al, 2018 ; Ashok et al, 2019 ; Mostafa et al, 2019 ; Safary et al, 2019 ; Karla et al, 2020 ; Prakash et al, 2020 ; Saeed et al, 2020 ; Prihanto et al, 2022 ). Further studies are necessary because there is still controversy regarding the coactivity of L-glutaminase asparaginase.…”

Diversity and bioactive profile of endophytic mycoflora in mangroves

“…Puri cation, characterization and antineoplastic behaviour of L-asparaginase from different endophytic fungi were studied (Manasa, 2014;Hatamazadeh, 2020;Moubasher, 2022). Trichoderma sp., a potent producer of l-asparaginase was isolated from Sonneratia alba (Prihanto, 2019). But the reports on enzyme, glutaminase from the endophytes is limited.…”