Zambia Mineral-Waste Bioremediation Predictor Metallothionein (MT) Computational Progress Report Author: Elsa Muleya | Institution: Copperbelt University / HTGAA External Cohort
Date: April 2026 | Project Phase: Aim 1 — Protein Identification & Construct Design
Table of Contents Project Overview Results Summary Detailed Results & Evidence 3.1 NCBI Protein Database Search 3.2 Chosen Protein Sequence 3.3 BLASTP Clustered NR Analysis 3.4 PHI-BLAST Analysis 3.5 Biochemical Properties (Benchling) 3.6 AlphaFold3 Structure Prediction 3.7 3D Structure Visualization 3.8 Construct Assembly in Benchling 3.9 pHT01 Backbone Verification (BLASTN) 3.10 Codon Optimization & Twist Order Computational Checklist Wet Lab Checklist References 1. Project Overview The Zambia Mineral-Waste Bioremediation Predictor aims to engineer Bacillus subtilis to express a heterologous metallothionein (MT) protein for the biosequestration of heavy metals (Cu²⁺, Co²⁺, Pb²⁺, Zn²⁺) contaminating water sources in the Copperbelt Province of Zambia. The system incorporates: