Type S Non-Ribosomal Peptide Synthetases for the Rapid Generation of Tailormade Peptide Libraries**

Bacterial natural products in general, and non-ribosomally synthesized peptides in particular, are structurally diverse and provide us with a broad range of pharmaceutically relevant bioactivities. Yet, traditional natural product research suffers from rediscovering the same scaffolds and has been stigmatized as inefficient, time-, labour- and cost-intensive. Combinatorial chemistry, on the other hand, can produce new molecules in greater numbers, cheaper and in less time than traditional natural product discovery, but also fails to meet current medical needs due to the limited biologically relevant chemical space that can be addressed. Consequently, methods for the high throughput generation of new natural products would offer a new approach to identifying novel bioactive chemical entities for the hit to lead phase of drug discovery programs. As a follow-up to our previously published proof-of-principle study on generating bipartite type S non-ribosomal peptide synthetases (NRPSs), we now envisaged the de novo generation of non-ribosomal peptides (NRPs) on an unreached scale. Using synthetic zippers, we split NRPSs in up to three subunits and rapidly generated different bi- and tripartite NRPS libraries to produce 49 peptides, peptide derivatives, and de novo peptides at good titres up to 145 mg L⁻¹. A further advantage of type S NRPSs not only is the possibility to easily expand the created libraries by re-using previously created type S NRPS, but that functions of individual domains as well as domain-domain interactions can be studied and assigned rapidly.     

Hardy和Littlewood的一个三角和

本文对Hardy和Littlewood考虑的一个有限三角和做了进一步地研究.通过充分运用Chebyshev多项式和M?bius函数的性质,建立了该有限三角和的一个有趣的恒等式,并得到了一个精确的渐近公式.     

Applications of Markov spectra for the weighted partition network by the substitution rule

The weighted self-similar network is introduced in an iterative way. In order to understand the topological properties of the self-similar network, we have done a lot of research in this field.Firstly, according to the symmetry feature of the self-similar network, we deduce the recursive relationship of its eigenvalues at two successive generations of the transition-weighted matrix.Then, we obtain eigenvalues of the Laplacian matrix from these two successive generations.Finally, we calculate an accurate expression for the eigentime identity and Kirchhoff index from the spectrum of the Laplacian matrix.     

Fq-linear codes over Fq-algebras

Huffman (2013) [12] studied ${\mathbb{F}}_q$-linear codes over ${\mathbb{F}}_{q^m}$ and he proved the MacWilliams identity for these codes with respect to ordinary and Hermitian trace inner products. Let S be a finite commutative ${\mathbb{F}}_q$-algebra. An ${\mathbb{F}}_q$-linear code over S of length n is an ${\mathbb{F}}_q$-submodule of $S^n$. In this paper, we study ${\mathbb{F}}_q$-linear codes over S. We obtain some bounds on minimum distance of these codes, and some large classes of MDR codes are introduced. We generalize the ordinary and Hermitian trace products over ${\mathbb{F}}_q$-algebras and we prove the MacWilliams identity with respect to the generalized form. In particular, we obtain Huffman's results on the MacWilliams identity. Among other results, we give a theory to construct a class of quantum codes and the structure of ${\mathbb{F}}_q$-linear codes over finite commutative graded ${\mathbb{F}}_q$-algebras.     

Multi-variable special polynomials using an operator ordering method

Using an operator ordering method for some commutative superposition operators, we introduce two new multi-variable special polynomials and their generating functions, and present some new operator identities and integral formulas involving the two special polynomials. Instead of calculating complicated partial differential, we use the special polynomials and their generating functions to concisely address the normalization, photocount distributions and Wigner distributions of several quantum states that can be realized physically, the results of which provide real convenience for further investigating the properties and applications of these states.     

Projective well orders and coanalytic witnesses

We further develop a forcing notion known as Coding with Perfect Trees and show that this poset preserves, in a strong sense, definable P-points, definable tight MAD families and definable selective independent families. As a result, we obtain a model in which $\mathfrak{a}=\mathfrak{u}=\mathfrak{i}={\mathrm{?}}_1<2^{{\mathrm{?}}_0}={\mathrm{?}}_2$, each of $\mathfrak{a}$, $\mathfrak{u}$, $\mathfrak{i}$ has a ${\mathrm{\Pi }}_1^1$ witness and there is a ${\mathrm{\Delta }}_3^1$ well-order of the reals. Note that both the complexity of the witnesses of the above combinatorial cardinal characteristics, as well as the complexity of the well-order are optimal. In addition, we show that the existence of a ${\mathrm{\Delta }}_3^1$ well-order of the reals is consistent with $\mathfrak{c}={\mathrm{?}}_2$ and each of the following: $\mathfrak{a}=\mathfrak{u}<\mathfrak{i}$, $\mathfrak{a}=\mathfrak{i}<\mathfrak{u}$, $\mathfrak{a}<\mathfrak{u}=\mathfrak{i}$, where the smaller cardinal characteristics have co-analytic witnesses.Our methods allow the preservation of only sufficiently definable witnesses, which significantly differs from other preservation results of this type.     

Development of a highly sensitive imaged cIEF immunoassay for studying AAV capsid protein charge heterogeneity

Post-translational modifications (PTMs) of adeno-associated virus (AAV) capsid proteins tune and regulate the AAV infective life cycle, which can impact the safety and efficacy of AAV gene therapy products. Many of these PTMs induce changes in protein charge heterogeneity, including deamidation, oxidation, glycation, and glycosylation. To characterize the charge heterogeneity of a protein, imaged capillary isoelectric focusing (icIEF) has become the gold standard method. We have previously reported an icIEF method with native fluorescence detection for denatured AAV capsid protein charge heterogeneity analysis. Although well suited for final products, the method does not have sufficient sensitivity for upstream, low-concentration AAV samples, and lacks the specificity for capsid protein detection in complex samples like cell culture supernatants and cell lysates. In contrast, the combination of icIEF, protein capture, and immunodetection affords significantly higher sensitivity and specificity, addressing the challenges of the icIEF method. By leveraging different primary antibodies, the icIEF immunoassay provides additional selectivity and affords a detailed characterization of individual AAV capsid proteins. In this study, we describe an icIEF immunoassay method for AAV analysis that is 90 times more sensitive than native fluorescence icIEF. This icIEF immunoassay provides AAV stability monitoring, where changes in individual capsid protein charge heterogeneity can be observed in response to heat stress. When applied to different AAV serotypes, this method also provides serotype identity with reproducible quantification of VP protein peak areas and apparent isoelectric point (pI). Overall, the described icIEF immunoassay is a sensitive, reproducible, quantitative, specific, and selective tool that can be used across the AAV biomanufacturing process, especially in upstream process development where complex sample types are often encountered.     

Combinatorial Screening of Water/Proton Permeation of Self-Assembled Pillar[5]arene Artificial Water Channel Libraries

Artificial water channels (AWCs) that selectively transport water and reject ions through bilayer membranes have potential to act as synthetic Aquaporins (AQPs). AWCs can have a similar osmotic permeability, better stability, with simpler manufacture on a larger-scale and have higher functional density and surface permeability when inserted into the membrane. Here, we report the screening of combinatorial libraries of symmetrical and unsymmetrical rim-functionalized PAs A – D that are able to transport ca. 10⁷–10⁸ water molecules/s/channel, which is within 1 order of magnitude of AQPs’ and show total ion and proton rejection. Among the four channels, C and D are 3–4 times more water permeable than A and B when inserted in bilayer membranes. The binary combinations of A – D with different molar ratios could be expressed as an independent (linear ABA ), a recessive (inhibition AB , AC , DB , ACA ), or a dominant (amplification, DBD ) behavior of the water net permeation events.