Towards more Sustainable Peptide- based Antibiotics: Stable in Human Blood, Enzymatically Hydrolyzed in Wastewater?

The emergence and spread of antibiotic resistance is a major societal challenge and new antibiotics are needed to successfully fight bacterial infections. Because the release of antibiotics into wastewater and downstream environments is expected to contribute to the problem of antibiotic resistance, it would be beneficial to consider the environmental fate of antibiotics in the development of novel antibiotics. In this article, we discuss the possibility of designing peptide-based antibiotics that are stable during treatment (e.g. in human blood), but rapidly inactivated through hydrolysis by peptidases after their secretion into wastewater. In the first part, we review studies on the biotransformation of peptide-based antibiotics during biological wastewater treatment and on the specificity of dissolved extracellular peptidases derived from wastewater. In the second part, we present first results of our endeavour to identify peptide bonds that are stable in human blood plasma and susceptible to hydrolysis by the industrially produced peptidase Subtilisin A

Towards more Sustainable Peptide- based Antibiotics: Stable in Human Blood, Enzymatically Hydrolyzed in Wastewater?

The emergence and spread of antibiotic resistance is a major societal challenge and new antibiotics are needed to successfully fight bacterial infections. Because the release of antibiotics into wastewater and downstream environments is expected to contribute to the problem of antibiotic resistance, it would be beneficial to consider the environmental fate of antibiotics in the development of novel antibiotics. In this article, we discuss the possibility of designing peptide-based antibiotics that are stable during treatment (e.g. in human blood), but rapidly inactivated through hydrolysis by peptidases after their secretion into wastewater. In the first part, we review studies on the biotransformation of peptide-based antibiotics during biological wastewater treatment and on the specificity of dissolved extracellular peptidases derived from wastewater. In the second part, we present first results of our endeavour to identify peptide bonds that are stable in human blood plasma and susceptible to hydrolysis by the industrially produced peptidase Subtilisin A

S_DGE.FQ1.tar.gz

Compressed file containing Tomato SAM RNA-seq reads from BrAd-seq DGE libraries (S_DGE_A5.fastq, S_DGE_A6.fastq, S_DGE_B7.fastq, S_DGE_C7.fastq

L_DGE.FQ1.tar.gz

Compressed file containing Tomato leaf RNA-seq reads from BrAd-seq DGE libraries (L_DGE_B5.fastq, L_DGE_C6.fastq, L_DGE_D7.fastq

SHO.FQ.tar.gz

Compressed file containing Tomato RNA-seq reads from BrAD-seq shotgun (SHO) type strand-specific libraries (SHO_22.fastq, SHO_23.fastq, SHO_24.fastq)

S_HTR.FQ.tar.gz

Compressed file containing Tomato SAM RNA-seq reads from HTR-method libraries (S_HTR_A5.fastq, S_HTR_A6.fastq, S_HTR_B6.fastq, S_HTR_B7.fastq

S_DGE.FQ2.tar.gz

Compressed file containing Tomato SAM RNA-seq reads from BrAd-seq DGE libraries (S_DGE_D8.fastq, S_DGE_E1.fastq, S_DGE_F2.fastq

L_DGE.FQ2.tar.gz

Compressed file containing Tomato leaf RNA-seq reads from BrAd-seq DGE libraries (L_DGE_E7.fastq, L_DGE_E8.fastq, L_DGE_F1.fastq

BrAD-seq: Breath Adapter Directional sequencing: a streamlined, ultra-simple and fast library preparation protocol for strand specific mRNA library construction.

Next Generation Sequencing (NGS) is driving rapid advancement in biological understanding and RNA-sequencing (RNA-seq) has become an indispensable tool for biology and medicine. There is a growing need for access to these technologies although preparation of NGS libraries remains a bottleneck to wider adoption. Here we report a novel method for the production of strand specific RNA-seq libraries utilizing the terminal breathing of double-stranded cDNA to capture and incorporate a sequencing adapter. Breath Adapter Directional sequencing (BrAD-seq) reduces sample handling and requires far fewer enzymatic steps than most available methods to produce high quality strand-specific RNA-seq libraries. The method we present is optimized for 3-prime Digital Gene Expression (DGE) libraries and can easily extend to full transcript coverage shotgun (SHO) type strand-specific libraries and is modularized to accommodate a diversity of RNA and DNA input materials. BrAD-seq offers a highly streamlined and inexpensive option for RNA-seq libraries