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Een added. Each point represents the mean and standard deviation of three independent experimentsproved the transcription of the three crystal proteins. Three other protoxins (Cry1F, insecticidal protein 2, and Cry1 type crystal protein) were also detected by 2D-LC?MS/MS in Cu medium. Cu2+ may activate the expression of specific protoxins. However, as theses protoxins determined had only one unique peptide detected, their expressions require further confirmation.Functional classificationThe proteins were classified into seven functional groups: small molecular metabolism, macromolecular metabolism, cell processes, regulation, environmental information processing proteins, extrachromosomal proteins, and unknown function (Fig. 7a; Additional files 2, 3, 4). Of these groups, the proteins were mainly distributed in small molecular metabolism (CK 39.32 , Cu 39.40 )and macromolecular metabolism (CK 24.42 , Cu 24.73 ). The orders of these categories showed no difference between the two media. Fold changes of functional categories between Cu and CK were computed (Fig. 8a), and the calculation was done as follows: the percentage of a specific category from Cu was divided by the percentage of that category from CK. A fold-change value >2 indicates a significant increase of a particular category in Cu, whereas a foldchange value Capecitabine after 46 h of cultivation in a the original medium and b the medium to which 10-6 mol/L Cu2+ had been addedHuang et al. [10] analyzed the proteome of B. thuringiensis subsp. kurstaki 4.0718 strain at different growth phases and category of proteins shows the lowest expression in the early sporulation phase, during which the synthesis of ICP was extremely active. Wang et al. [11] found that most genes involved in amino acid transport and biosynthesis are downregulated in the sporulationperiod, which indicates that the amino acid sources from these two pathways are limited. Thus, that amino acid requirements during sporulation are fulfilled by PubMed ID:https://www.ncbi.nlm.nih.gov/pubmed/16989806 protein turnover was proposed. Monro [12] proved that 80 of the amino acids for ICP synthesis come from protein turnover through a radioisotopic tracer experiment. Protein turnover as the major source of amino acids duringLiu et al. Microb Cell Fact (2015) 14:Page 7 ofTable 1 LC50 analysis of B. thuringiensis X022 fermentation broth from two different mediumMedium CKa Cub LC50 (g/mL) to S. exigua 1.052 0.484 95 CI 0.692?.172 0.257?.544 Slope 0.085 0.073 LC50 (g/mL) to H. armigera 0.518 0.240 95 CI 0.371?.864 0.146?.441 Slope 0.075 0.Non-overlapping 95 confidence intervals of LC50 were used as the criteria to determine significant difference of toxicities among different formulations of acetamiprid CI confidence intervala bCK represent fermentation broth from the original medium Cu represent fermentation broth from 10-6 mol/L Cu2+ added medi.