Journal: Applied Microbiology and Biotechnology

Article Title: Heterologous expressing melittin in a probiotic yeast to evaluate its function for promoting NSC-34 regeneration

doi: 10.1007/s00253-024-13336-7

Figure Lengend Snippet: MET and ProMET construction and Western blotting assays in a yeast culture supernatant. A Schematic representation of K. marxianus e expressing MET and ProMET ; B Western blotting analysis of MET; C Western blotting analysis of ProMET. Lane 1, day 1; lane 2, day 2; lane 3, day 3; lane 4, day 4

Article Snippet: MET and precursor ProMET genes from Apis mellifera were codon-optimized toward K. marxianus and synthesized (Twist Bioscience, USA), which sequences are shown in Fig. . Sequence design and optimization for the two constructions (pKLAC2-a-his-MET and pKLAC2-a-his-ProMET) were inserted in a plasmid (pKLAC2-a-) between Nde I and Not I restriction enzyme sites.

Techniques: Western Blot, Expressing

Journal: Applied Microbiology and Biotechnology

Article Title: Heterologous expressing melittin in a probiotic yeast to evaluate its function for promoting NSC-34 regeneration

doi: 10.1007/s00253-024-13336-7

Figure Lengend Snippet: MET and ProMET purification analysis through histidine-affinity chromatography and desalting columns. A MET; B ProMET. Lane 1, flow-through; lane 2, wash; lane 3, elution; lane 4, desalting flow-through; lane 5, desalting elution

Article Snippet: MET and precursor ProMET genes from Apis mellifera were codon-optimized toward K. marxianus and synthesized (Twist Bioscience, USA), which sequences are shown in Fig. . Sequence design and optimization for the two constructions (pKLAC2-a-his-MET and pKLAC2-a-his-ProMET) were inserted in a plasmid (pKLAC2-a-) between Nde I and Not I restriction enzyme sites.

Techniques: Purification, Affinity Chromatography

Journal: Applied Microbiology and Biotechnology

Article Title: Heterologous expressing melittin in a probiotic yeast to evaluate its function for promoting NSC-34 regeneration

doi: 10.1007/s00253-024-13336-7

Figure Lengend Snippet: Identified peptide profiles of purified MET and ProMET compared with natural and commercial crude BV extract through HPLC. *BV-A4-1, natural crude BV extract; BV-CITEQ, commercial crude BV extract; MET, purified MET from this study; ProMET, purified ProMET from this study; MET-no TEV, TEV protease untreated MET. **Peak numbers: I, melittin or promelittin major peak; II, an unknown compound; III, non-protease treated MET constructed in this study

Article Snippet: MET and precursor ProMET genes from Apis mellifera were codon-optimized toward K. marxianus and synthesized (Twist Bioscience, USA), which sequences are shown in Fig. . Sequence design and optimization for the two constructions (pKLAC2-a-his-MET and pKLAC2-a-his-ProMET) were inserted in a plasmid (pKLAC2-a-) between Nde I and Not I restriction enzyme sites.

Techniques: Purification, Construct

Journal: Applied Microbiology and Biotechnology

Article Title: Heterologous expressing melittin in a probiotic yeast to evaluate its function for promoting NSC-34 regeneration

doi: 10.1007/s00253-024-13336-7

Figure Lengend Snippet: MS/MS spectrum of A MET and B ProMET sequences

Article Snippet: MET and precursor ProMET genes from Apis mellifera were codon-optimized toward K. marxianus and synthesized (Twist Bioscience, USA), which sequences are shown in Fig. . Sequence design and optimization for the two constructions (pKLAC2-a-his-MET and pKLAC2-a-his-ProMET) were inserted in a plasmid (pKLAC2-a-) between Nde I and Not I restriction enzyme sites.

Techniques: Tandem Mass Spectroscopy

Journal: Applied Microbiology and Biotechnology

Article Title: Heterologous expressing melittin in a probiotic yeast to evaluate its function for promoting NSC-34 regeneration

doi: 10.1007/s00253-024-13336-7

Figure Lengend Snippet: Safety assessment of purified MET and ProMET from treating NSC-34 neuron cells through the cell counting Kit-8. A Standard curve; B cell viability. *BV-A4-1, natural crude BV extract; MET, purified MET from this study; ProMET, purified ProMET from this study. **Values are averages of three replicated analyses with standard deviations. Significant differences ( p < 0.05) between groups are indicated by the different letters above the bars

Article Snippet: MET and precursor ProMET genes from Apis mellifera were codon-optimized toward K. marxianus and synthesized (Twist Bioscience, USA), which sequences are shown in Fig. . Sequence design and optimization for the two constructions (pKLAC2-a-his-MET and pKLAC2-a-his-ProMET) were inserted in a plasmid (pKLAC2-a-) between Nde I and Not I restriction enzyme sites.

Techniques: Purification, Cell Counting

Journal: Applied Microbiology and Biotechnology

Article Title: Heterologous expressing melittin in a probiotic yeast to evaluate its function for promoting NSC-34 regeneration

doi: 10.1007/s00253-024-13336-7

Figure Lengend Snippet: NSC-34 neuron cell regeneration fold change following H 2 O 2 damage and treated with groups. A 12 h; B 24 h; C 48 h. *BV-A4-1, natural crude BV extract; BV-CITEQ, commercial crude BV extract; MET, purified MET from this study; ProMET, purified ProMET from this study; Control, non-H 2 O 2 -damaged control. **Values are averages of three replicated analyses with standard deviations. Significant differences ( p < 0.05) between groups are indicated by the different letters above the bars

Article Snippet: MET and precursor ProMET genes from Apis mellifera were codon-optimized toward K. marxianus and synthesized (Twist Bioscience, USA), which sequences are shown in Fig. . Sequence design and optimization for the two constructions (pKLAC2-a-his-MET and pKLAC2-a-his-ProMET) were inserted in a plasmid (pKLAC2-a-) between Nde I and Not I restriction enzyme sites.

Techniques: Purification, Control

Journal: Applied Microbiology and Biotechnology

Article Title: Heterologous expressing melittin in a probiotic yeast to evaluate its function for promoting NSC-34 regeneration

doi: 10.1007/s00253-024-13336-7

Figure Lengend Snippet: Gene expression analysis of growth factors in NSC-34 neuron cells treated with groups through qPCR. *BV-A4-1, natural crude BV extract; MET, purified MET from this study; ProMET, purified ProMET from this study. **Values are averages of three replicated analyses with standard deviations. Significant differences ( p < 0.05) between groups are indicated by the different letters above the bars

Article Snippet: MET and precursor ProMET genes from Apis mellifera were codon-optimized toward K. marxianus and synthesized (Twist Bioscience, USA), which sequences are shown in Fig. . Sequence design and optimization for the two constructions (pKLAC2-a-his-MET and pKLAC2-a-his-ProMET) were inserted in a plasmid (pKLAC2-a-) between Nde I and Not I restriction enzyme sites.

Techniques: Gene Expression, Purification