J. Pharm. Pharmacogn. Res., vol. 14, no. 1, Article ID 2340, Jan-Feb 2026.

DOI: https://doi.org/10.56499/jppres_14.1.2340

Original Article

Isolation and characterization of Bacillus subtilis BPP16: A marine-derived α-amylase producer

[Aislamiento y caracterización de Bacillus subtilis BPP16: un productor de α-amilasa de origen marino]

Mochamad Fathurohman^1,2, Tursino¹, Muhammad Azhari¹, Anjali Patel¹, Elin Julianti^1*

¹Department of Pharmacochemistry, School of Pharmacy, Insitut Teknologi Bandung, 40132 Bandung, Indonesia.

²Departement of Pharmacochemistry, Faculty of Pharmacy, Bakti Tunas Husada University, 46115 Tasikmalaya, Indonesia.

^*E-mail: elin_julianti@itb.ac.id

Abstract

Context: α-Amylase is an essential industrial enzyme widely applied in the conversion of starch into simple sugars. Marine environments are a promising source of novel enzyme-producing microorganisms, owing to their diverse and adaptive microbial communities.

Aims: To isolate and characterize amylolytic microorganisms from marine sources and to evaluate the α-amylase activity of selected isolates.

Methods: Samples were collected from the marine zone of Pramuka Island, Indonesia. Isolation of bacteria and fungi was performed using modified Zobell media, followed by screening for α-amylase activity using starch agar and iodine staining. The most potent isolate, BPP16, underwent morphological, biochemical, and molecular characterization via 16S rRNA gene analysis. Crude α-amylase was extracted through liquid-state fermentation and its activity quantified using the dinitrosalicylic acid (DNS) assay.

Results: A total of 27 pure microbial cultures (19 bacterial, 8 fungal) were obtained, with 18 bacterial isolates demonstrating α-amylase activity. Isolate BPP16 exhibited the highest amylolytic index (5.23 ± 0.07). Characterization revealed Gram-positive, rod-shaped morphology consistent with Bacillus subtilis. Biochemical assays indicated positive results for indole, methyl red, Voges-Proskauer, catalase, and motility tests, with glucose fermentation observed. Molecular analysis confirmed 100% sequence identity with Bacillus subtilis. The crude enzyme showed an activity of 10.29 IU/mg and a specific activity of 3.62 U/mg.

Conclusions: The marine-derived Bacillus subtilis BPP16 exhibited substantial α-amylase activity, suggesting its potential for industrial applications in starch processing.

Keywords: α-amylase; Bacillus subtilis BPP16; enzyme activity; industrial biotechnology; marine microorganisms.

Resumen

Contexto: La α-amilasa es una enzima industrial esencial, ampliamente utilizada para convertir el almidón en azúcares simples. Los ambientes marinos constituyen una fuente prometedora de microorganismos capaces de producir nuevas enzimas, debido a la diversidad y la adaptabilidad de sus comunidades microbianas.

Objetivos: Aislar y caracterizar microorganismos amilolíticos de origen marino y evaluar la actividad de la α-amilasa en aislados seleccionados.

Métodos: Se recolectaron muestras en la zona marina de la isla de Pramuka, Indonesia. El aislamiento de bacterias y hongos se realizó mediante un medio de Zobell modificado, seguido de la detección de la actividad de la α-amilasa mediante agar almidón y tinción con yodo. El aislado más potente, BPP16, se sometió a caracterización morfológica, bioquímica y molecular mediante el análisis del gen ARNr 16S. La α-amilasa cruda se extrajo mediante fermentación en estado líquido y su actividad se cuantificó mediante el ensayo de DNS.

Resultados: Se obtuvieron 27 cultivos microbianos puros (19 bacterianos y 8 fúngicos), de los cuales 18 aislados bacterianos mostraron actividad α-amilasa. El aislado BPP16 presentó el índice amilolítico más alto (5,23 ± 0,07). La caracterización reveló una morfología grampositiva, en forma de bastón, consistente con Bacillus subtilis. Los ensayos bioquímicos indicaron resultados positivos para las pruebas de indol, rojo de metilo, Voges-Proskauer, catalasa y motilidad, observándose fermentación de glucosa. El análisis molecular confirmó una identidad de secuencia del 100 % con Bacillus subtilis. La enzima cruda mostró una actividad de 10,29 UI/mg y una actividad específica de 3,62 U/mg.

Conclusiones: El Bacillus subtilis BPP16, de origen marino, exhibió una actividad α-amilasa sustancial, lo que sugiere su potencial para aplicaciones industriales en el procesamiento del almidón.

Palabras Clave: actividad enzimática; α-amilasa; Bacillus subtilis BPP16; biotecnología industrial; microorganismos marinos.

 

https://jppres.com/jppres/pdf/vol14/jppres_14.1.2340.pdf

 

 

Citation Format: Fathurohman M, Tursino, Azhari M, Patel A, Julianti E (2026) Isolation and characterization of Bacillus subtilis BPP16: A marine-derived α-amylase producer. J Pharm Pharmacogn Res 14(1): 2340. https://doi.org/10.56499/jppres_14.1.2340

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