Application of scanning electron microscopy and MALDI-TOF MS in the exploration of the parasporal insecticidal crystals of Bacillus thuringiensis strains isolated from Qatar soil

Al-Thani, Aldana; Ashfaq, Mohammad Y.; Al‐Thani, Roda; Hassan, Zahoor Ul; Jaoua, Samir

doi:10.1016/j.biteb.2022.101134

Cited by 2 publications

(1 citation statement)

References 30 publications

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“…Bacillus thuringiensis is a type of gram-positive bacteria that has been widely used in the field of agricultural biotechnology because it has toxicity on several plant insect pests such as the order Diptera and Lepidoptera [1]. There are at least 70 different subspecies or varieties of B. thuringiensis known to produced more than 300 Cry proteins from 1000 strains and only a few strains have been utilized [2].…”

Section: Introductionmentioning

confidence: 99%

Sodium dodecyl sulphate polyacrylamide gel electrophoresis of toxin proteins of Bacillus thuringiensis isolated from soil at the Liwa Botanical Garden

Kusuma Handayani,

Muhammad Rian Hidayat,

Wawan Abdullah Setiawan

et al. 2023

GSC Biol. Pharm. Sci.

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Bacillus thuringiensis is a spore-forming bacterium known to have the ability to synthesize proteins that are toxic against insect pests. Currently, the bacterial protein toxins are widely used as pesticides because they are environmentally friendly and safe for health. As a cosmopolitan microbe B. thuringiensis is widely distributed in nature with soil as its main habitat. Liwa Botanical Gardens is one among the conservation areas in Lampung Province of Indonesia where the soil type and conditions expected to good for the bacterial life of B. thuringiensis. This study aims to determine toxin proteins extracted from B. thuringiensis isolated from the soil of the Liwa Botanical Garden by their molecular weights. The proteins were quantified using Lowry method and the protein profiling performed using SDS PAGE electrophoresis. The results showed that there were two isolates, Bt 3 and Bt 5 which showed close protein concentrations, namely 0.679 mg/ml and Bt 5 with 1.313 mg/ml respectively. Both isolates also exhibit different molecular weights. Bt 3 isolate has a molecular weight of 110.59 kDa, while Bt 5 has a molecular weight of 76.97 kDa.

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Section: Introductionmentioning

confidence: 99%

Sodium dodecyl sulphate polyacrylamide gel electrophoresis of toxin proteins of Bacillus thuringiensis isolated from soil at the Liwa Botanical Garden

Kusuma Handayani,

Muhammad Rian Hidayat,

Wawan Abdullah Setiawan

et al. 2023

GSC Biol. Pharm. Sci.

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A dye elution method for the quantification of insecticidal crystal proteins from Bacillus thuringiensis and its compatibility with the presence of agro-industrial raw materials and waste products

Mentel,

Loto,

Baigori

et al. 2024

Letters in Applied Microbiology

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The insecticidal crystal proteins produced by Bacillus thuringiensis during sporulation are active ingredients against lepidopteran, dipteran, and coleopteran insects. Several methods have been reported for their quantification such as crystals counting, ELISA, and SDS-PAGE/densitometry. One of the major tasks in industrial processes is the analysis of raw materials dependency and costs. Thus, the crystal proteins quantification method is expected to be compatible with the presence of complex and inexpensive culture medium components. This work presents a revalidated elution-based method for the quantification of insecticidal crystal proteins produced by the native strain B. thuringiensis RT. To quantify proteins, a calibration curve was generated by varying the amount of BSA loaded into SDS-PAGE gels. Firstly, SDS-PAGE was performed for quality control of the bioinsecticide. Then, the stained protein band was excised from 10% polyacrylamide gel and the protein-associated dye was eluted with an alcoholic solution of SDS (3% SDS in 50% isopropanol) during 45 min at 95°C. This protocol was a sensitive procedure to quantify proteins in the range of 2.0–10.0 µg. As proof of concept, proteins of samples obtained from a complex fermented broth were separated by SDS-PAGE. Then, Cry1 and Cry2 proteins were properly quantified.

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Application of scanning electron microscopy and MALDI-TOF MS in the exploration of the parasporal insecticidal crystals of Bacillus thuringiensis strains isolated from Qatar soil

Cited by 2 publications

References 30 publications

Sodium dodecyl sulphate polyacrylamide gel electrophoresis of toxin proteins of Bacillus thuringiensis isolated from soil at the Liwa Botanical Garden

Sodium dodecyl sulphate polyacrylamide gel electrophoresis of toxin proteins of Bacillus thuringiensis isolated from soil at the Liwa Botanical Garden

A dye elution method for the quantification of insecticidal crystal proteins from Bacillus thuringiensis and its compatibility with the presence of agro-industrial raw materials and waste products

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