Les poly(3-hydroxyalcanoate)s ou PHAs sont des biopolyesters linéaires biodégradables et biocompatibles synthétisés par des microorganismes bactériens en tant que réserve de carbone et d'énergie. Ils sont synthétisés par des bactéries à partir de ressources renouvelables et la diversité de leurs structures possibles se traduit par un large éventail de polymères ayant des propriétés mécaniques très différentes. Nous avons tout d'abord testé les capacités de production de PHAs de nouvelles souches bactériennes marines provenant de tapis microbiens de Polynésie française, en utilisant, entre autres, des substrats naturels comme l'huile de coprah, le glucose et l'acide oléique. Nous avons notamment montré que la souche Pseudomonas guezennei est capable de produire des PHAs avec des taux d'insaturation contrôlés et de masse molaire très élevée. Puis, des oligomères de PHAs fonctionnalisés de structures contrôlées portant des fonctions terminales alcynes ou alcènes ont été préparés par transestérification. Ces oligomères ont ensuite été utilisés pour l'élaboration par chimie click de copolymères amphiphiles greffés EPS-g-PHA avec des exopolysaccharides (EPS) bactériens. Enfin la dernière partie de ces travaux a consisté en la réalisation d'un support de croissance pour le développement de cellules souches pour l'ingénierie tissulaire combinant les propriétés mécaniques des PHAs et les propriétés hydrophiles et bioactives des EPS

This study has researched how user click heatmaps and eye tracking can be utilized in usability testing. We have used the tools SessionCam and Tobii. We designed a website for a small clothing store and conducted usability tests, in order to evaluate the users clicks and eye movements in relation to design choices on the website. The aim of this study was to research how these tools can be integrated in usability testing and how results from the tests can be used to improve usability. We were able to use the recordings of clicks and eye movements and make design changes based on the findings. Some of the findings were that the users look more at the top of the page than at the bottom. They also look more at icons and images rather than longer text blocks. From the tracking of user clicks we found that the users primarily clicked on links in order to navigate the page but also that they sometimes clicked on texts when they were reading.

Two classes of ion-channels comprising 22 members were prepared. Three members were linear oligo-esters with terephthalate core designed to span both leaflets of the bilayer; these were prepared in a modular synthesis in three linear steps. 19 half-channels based on cyclodextrins with functionalized primary-rims were prepared by the Huisgen Cu+-catalyzed [3+2]-cyclization; three distinct synthetic protocols were established to be applicable to these substrates. The voltage-clamp experiment was used to characterize the ion transport properties of these 22 compounds as well as 5 oligo-esters previously prepared by solid-phase synthesis. All but two were active in bilayers, with the majority of these compounds showing highly complex conductance activities. Exponentially voltage-dependent currents were observed for two compounds (both terephthalate-derived); exclusive “square-top” activities were observed for one solid-phase–derived compound and one cyclodextrin-based channels; fractal openings were observed for at least two cyclodextrin-based channels. An “activity grid” notation was proposed as an empirical, coarse, but model-free method of treating the complex data. Through an exhaustive analysis of previously published synthetic ion channels, disparate compounds were found to share modes of activity. Supporting software were developed to facilitate the preparation of activity grids from current traces acquired for the aforementioned 27 compounds. Resulting activity grids for individual experiments were collated to generate an activity profile for each compound, from which a structure–activity map was established and could be compared to the literature data. Four core findings emerged. First, the activity grid notation is sufficiently expressive to denote highly complex mixture of activities. Second, systematic application of the notation reduces selection bias in data analysis. Third, many synthetic ion channels share highly sim- ilar activities and suggests the participation of the lipids, water, and ions in pore-formation. Lastly, the cyclodextrin half-channels are generally membrane active, and their activities are clearly modulated by structural variations. / Graduate

Histone deacetylase (HDAC) inhibition is an emerging novel therapeutic strategy in cancer therapy. HDAC inhibitors (HDACi) have shown ability to block angiogenesis and cell cycling, as well as initiate differentiation and apoptosis. In fact, suberoylanilide hydroxamic acid (SAHA) is the first in the class of HDACi approved by the FDA for the treatment of cutaneous T cell lymphoma. On the other hand, there is a sustained interest in the use of gold nanoparticles (AuNPs) for various cancer diagnostic and therapeutic applications - bioimaging, drug delivery, and binary therapy techniques such as photodynamic and photothermal therapies. This interest in AuNPs is facilitated by favorable attributes such as ease of fabrication, bioconjugation and biocompatibility, and unique optical and electronic properties. However, HDACi- and AuNPs- based antitumor agents are plagued with problems common to all chemotherapeutic agents such as lack of selectivity, which often results in systemic toxicity. Therefore, availability of a methodology to selectively deliver AuNPs and HDACi to cancer cells will significantly improve their therapeutic indices and lead to the identification of novel agents for use in diagnostic imaging and targeted cancer therapy applications.

A Internet vem transformando a maneira como as pessoas compram e buscam por informações sobre produtos. Para locais onde as ofertas físicas apresentam algum tipo de restrição, os canais online podem trazer mudanças ainda mais significativas, já que oferecem aos consumidores um sortimento virtualmente ilimitado. O objetivo deste trabalho foi o de analisar a influência do número de habitantes de uma cidade em relação à predisposição à seleção e compra de produtos de uma empresa que administra campanhas de divulgação pela Internet para venda por atacado. As cidades foram divididas em três grupos conforme o número de habitantes e as principais métricas de performance das campanhas online foram comparadas entre os grupos através de análises de estatísticas descritivas, ANOVA e testes post hoc. Os grupos apresentaram diferenças estatisticamente significativas entre si em todas as etapas do processo de busca e compra. A principal diferença encontrada foi no momento em que os usuários preencheram cadastros no site, demonstrando interesse nos produtos. Nesta etapa, os consumidores de municípios de menor porte se cadastraram proporcionalmente mais do que o dobro dos de cidades de porte intermediário e mais de três vezes em relação ao grupo das maiores, o que impactou diretamente no custo por venda realizada e nas taxas absolutas de vendas. / The Internet has been changing the way people buy and search for new products. In places where the assortment in brick-and-mortar stores is restricted, online channels may represent a significant change in the landscape, due to the fact that they carry virtually unlimited offers. The aim of this study is to analyze the influence of the number of inhabitants of a city on their likelihood of selecting and purchasing products from a company which runs online marketing campaigns for resellers. The cities were divided in three groups according to the population size. In addition, the main metrics for online campaigns were compared among groups through descriptive statistics, ANOVAs and pot hoc tests. There was a statistically significant difference among the groups in all steps of the search and purchase process. The main difference was found when the users fill in the forms in the website, demonstrating interest in the product. In this stage, customers from smaller cities showed a registering rate which was more than twice as much higher than the medium-sized cities group and more than three times the amount of the larger cities group, which led to a direct impact on the cost-per-sale and absolute sales rates.

Le développement de nouveaux systèmes pour la délivrance locale et contrôlée de principes actifs (PA) peut avoir plusieurs intérêts : augmenter la biodisponibilité du PA, délivrer une forte dose prolongée en limitant les effets secondaires, et améliorer le confort du patient. Pour développer ces nouveaux systèmes de délivrance, des matériaux composites ont été élaborés. Ces matériaux sont constitués d’une matrice hydrogel intégrant des nanoparticules lipidiques (LNPs) capables de solubiliser un principe actif hydrophobe. Cette double encapsulation a pour but de solubiliser puis de délivrer un principe actif hydrophobe qui ne pourrait pas être encapsulé directement dans la matrice hydrophile, et de retarder sa libération en créant des interactions particules/polymères. Nous avons élaboré différents matériaux en sélectionnant 2 polysaccharides largement utilisés pour des applications biomédicales : la carboxyméthylcellulose (CMC) et le chitosane (CS). Ces 2 polymères ont été mélangés à du poly(éthylène glycol) (PEG), un polymère synthétique flexible, pour augmenter les possibilités de moduler la structure et les propriétés mécaniques de la matrice. Ces hydrogels hybrides de CMC/PEG ou CS/PEG réticulés chimiquement par chimies clics, ont été proposés sous forme d’hydrogel en volume, de films ou d’éponges. Après un travail sur le développement et la caractérisation de ces nouvelles matrices hydrogels, les propriétés des matériaux composites polysaccharides/LNPs ont été évaluées et corrélées à la diffusion des particules dans le réseau hydrogel. / Due to their high biocompatibility, macroscale hydrogels have been studied as promising materials for the design of drug delivery systems (DDS). Such systems devoted to the local administration and prolonged drug release can improve the efficacy of pharmaceutical coumpounds while limiting undesired side-effects. Hydrogels present a high water content and soft consistency with mechanical properties that can match those of biological tissues. Nevertheless, these systems are essentially limited to the delivery of hydrophilic drugs. Our approach for extended release of hydrophobic drugs is to design composite materials composed of lipid nanoparticles (LNPs) entrapped within polysaccharide hydrogels. We selected two polysaccharides which are currently used in pharmaceutical and biomedical applications: carboxymethylcellulose (CMC) and chitosan (CS). We also used poly(ethylene glycol) (PEG) as a plasticizer to tune the matrix mechanical properties. Three types of LNP-loaded hybrid materials were studied; i) bulk CMC/PEG hydrogels, ii) CS/PEG films, and iii) CS/PEG sponges. These materials were chemically crosslinked through attractive click reactions. LNPs were successfully entrapped within the three materials without affecting their properties. A deeper study was conducted with the CMC/PEG composite hydrogel. The LNP release profiles were correlated with the network structure and particles properties. The different materials appear promising systems for the time-controlled delivery of therapeutics.

A Internet vem transformando a maneira como as pessoas compram e buscam por informações sobre produtos. Para locais onde as ofertas físicas apresentam algum tipo de restrição, os canais online podem trazer mudanças ainda mais significativas, já que oferecem aos consumidores um sortimento virtualmente ilimitado. O objetivo deste trabalho foi o de analisar a influência do número de habitantes de uma cidade em relação à predisposição à seleção e compra de produtos de uma empresa que administra campanhas de divulgação pela Internet para venda por atacado. As cidades foram divididas em três grupos conforme o número de habitantes e as principais métricas de performance das campanhas online foram comparadas entre os grupos através de análises de estatísticas descritivas, ANOVA e testes post hoc. Os grupos apresentaram diferenças estatisticamente significativas entre si em todas as etapas do processo de busca e compra. A principal diferença encontrada foi no momento em que os usuários preencheram cadastros no site, demonstrando interesse nos produtos. Nesta etapa, os consumidores de municípios de menor porte se cadastraram proporcionalmente mais do que o dobro dos de cidades de porte intermediário e mais de três vezes em relação ao grupo das maiores, o que impactou diretamente no custo por venda realizada e nas taxas absolutas de vendas. / The Internet has been changing the way people buy and search for new products. In places where the assortment in brick-and-mortar stores is restricted, online channels may represent a significant change in the landscape, due to the fact that they carry virtually unlimited offers. The aim of this study is to analyze the influence of the number of inhabitants of a city on their likelihood of selecting and purchasing products from a company which runs online marketing campaigns for resellers. The cities were divided in three groups according to the population size. In addition, the main metrics for online campaigns were compared among groups through descriptive statistics, ANOVAs and pot hoc tests. There was a statistically significant difference among the groups in all steps of the search and purchase process. The main difference was found when the users fill in the forms in the website, demonstrating interest in the product. In this stage, customers from smaller cities showed a registering rate which was more than twice as much higher than the medium-sized cities group and more than three times the amount of the larger cities group, which led to a direct impact on the cost-per-sale and absolute sales rates.

近年來行動網路及數位商務的蓬勃發展，促使O2O(Online to Offline/ Offline to Online)模式的興起。如何有效的結合實體通路與電子商務平台，以發展線上及線下之O2O模式，已成為提升產業競爭力的重要關鍵。 銷售商品不只透過實體店面進行販售，網路的普及化造就了網路銷售平台的架設，大型的購物平台對於零售業產生巨大的影響，因此企業不得不將實體及網路做整合。在傳統的電子商務，不外乎為 B2B、B2C、C2C 等型態，這些模式均屬於單方面虛擬店面的交易。隨著智慧型手機的普及，行動商務已是企業不可忽視的商機。 在現今的社會裡，網際網路帶動了電子商務的流行趨勢，也引導企業走向資訊化行銷的經營模式。利用虛實整合的策略，可以在真實世界與網路虛擬世界間，搭起一座溝通的橋樑，由傳統企業擴展電子商務的市場。在網際網路新世紀中，唯有把實體與虛擬作良好的整合，才能建立並維持競爭優勢。 當企業將實體及網路零售做整合時，實體店面增加了網路上的通路宣傳資源，方便追蹤消費者每筆消費紀錄，對企業來說能夠更輕鬆掌握消費者數據，可以提升對消費者的營銷效果及維護。本論文研究探討對於線上線下的整合，是否有利整體績效呈現，運用對O2O的文獻回顧探討線上線下的整合如何提升營運績效與競爭力。 / In recent years, the vigorous development of the action network and digital commerce has prompted the rise of the O2O model (Online to Offline/ Offline to Online). How to effectively combine physical access and e-commerce platform to develop the model O2O of online and offline has become an important key to enhance the competitiveness of the industry. Sales goods are there are nothing more than pattern sold through physical stores, but the popularization of Internet has created the network sales platform. Large shopping platforms have a huge impact on the retail industry, so enterprises have to integrate entities and networks. In the traditional e-commerce, not only for B2B, B2C, C2C, and so on, these patterns belong to the unilateral virtual store transaction. With the popularity of smart phone, mobile commerce is a business is a business opportunity to be reckoned with in enterprises. In today's society, the Internet has led to the trend of e-commerce, and also leads to the business model of information marketing. The strategy of virtual reality integration can set up a bridge of communication between the real world and the virtual world of the Internet, and expand the market of e-commerce by traditional enterprises. In the new century of the Internet, the competition advantage can be established and maintained only if the entity and the virtual are well integrated. When enterprises integrate entities and online retailers, the physical stores increase the channel publicity resources on the Internet, which is easy to track consumers' consumption records, and can easily grasp the consumer data, and enhance the effectiveness of consumer marketing and maintenance. This paper investigates whether the integration of online and offline whether beneficial to the overall performance or not, and uses literature reviews of O2O to explore how online-offline integration can improve operational performance and competitiveness.

La grande quantité des solvants utilisés dans l'industrie chimique représente un des problèmes les plus importants à aborder, d'un point de vue environnemental ainsi qu'économique. Les solvants communément utilisés sont volatiles, toxiques, inflammables et/ou corrosifs, et leur substitution par des autres moins nocifs est un objectif primordiale dans un cadre de développement durable. Le glycérol se présente comme un candidat prometteur pour remplacer les solvants organiques conventionnels. Il est généré comme produit concomitant dans la production de biodiesel. En conséquence, la valorisation du glycérol est devenue un enjeu majeur. Ce composé possède des propriétés très intéressantes pour être utilisé comme solvant en catalyse, telles que son innocuité, haut point d'ébullition, pression de vapeur négligeable, capacité de solubilisation de composés organiques et inorganiques, faible miscibilité avec autres solvants organiques et également son faible coût. Cette Thèse à pour objectif développer des nouveaux systèmes catalytiques dans un milieu glycérol. En particulier, dans ce travail nous montrons comme le glycérol promue la stabilisation de nanoparticules de cuivre (I) capables de catalyser la réaction de Huisgen 1,3-dipolaire entre alcynes terminaux et azotures organiques (connu comme la réaction click), de façon très efficace à température ambiante. Dans ce solvant, cette réaction peut aussi être réalisée en absence de cuivre avec alcynes internes non activés, en travaillant sous activation microondes. L'utilisation du glycérol favorise la réaction par rapport à des autres solvants (incluant ceux qui sont protiques), probablement grâce à sa capacité pour former liaisons d'hydrogène, favorisant l'interaction avec l'irradiation microondes (et ainsi accélérant la réaction). Avec le propos d'étudier des transformations stéréosélectives, nous avons préparé des nouveaux ligands énantiopures dérivés du PTA (1,3,5-triaza-7-phosphaadamantane). Ces composés innovants ont été utilisés en réactions énantiosélectives comme l'addition asymétrique de B2pin2 à N-Boc-imines (Cu) ou encore la alpha-amination (Cu), ou encore comme organocatalyseur (réaction de Morita-Baylis- Hillman). / Solvents are used in huge amounts in the chemical industry and this is one of the biggest problems to be solved not only from an environmental point of view, but also for economic reasons. The solvents commonly used in the industry are volatile, toxic, inflammable and/or corrosive; then, their replacement by others less harmful represents a crucial objective. Glycerol appears as a promising candidate to substitute the conventional organic solvents. It is produced in huge amounts as a waste in biodiesel industry. Consequently, the valorisation of glycerol becomes an important concern. This compound possesses very interesting properties to be used as solvent such as its non-toxicity, the wide range of temperature for its liquid state, negligible vapour pressure, capacity for solubilising organic and inorganic compounds, low miscibility with other organic solvents and also its low price. This Thesis deals with the development of new catalytic systems in glycerol medium. In particular, in this work we show how glycerol can facilitate the stabilisation of copper(I) nanoparticles that are able to catalyse the 1,3-dipolar Huisgen cycloaddition between terminal alkynes and organic azides (known as click reaction). The reaction proceeds at room temperature and it is very efficient. Moreover, this reaction can be carried out in the absence of copper using non-activated internal alkynes, working under microwave irradiation. Glycerol favours the process in comparison with other solvents (including protic ones), probably due to its ability to form hydrogen bonds, which favours the interaction with the microwave irradiation (accelerating the process). With the aim of studying stereoselective transformations, we have conceived new enantiopure ligands derived from PTA (1,3,5-triaza-7-phosphaadamantane). These innovative phosphines have been applied in enantioselective processes, such as pinacolboryl addition to N-Boc-imines (Cu) or alpha-amination (Cu), among others, or as organocatalyst (Morita-Baylis-Hillman reaction).

Le développement des technologies de recombinaison en biologie moléculaire fut un point tournant pour les sciences biologiques. Depuis cette découverte, diverses avancées extraordinaires qui ont un impact direct sur les humains ont pu être accomplies dans les domaines de recherches qui découlent de cette technologie. L’étude des enzymes produites en utilisant cette technique est le fondement de leurs applications éventuellement accessibles. À cet effet, la biocatalyse est un sous-domaine de l’enzymologie en développement continuel. Les chimistes et ingénieurs utilisent les composantes de systèmes biologiques ou même des systèmes complets afin de complémenter ou remplacer des méthodologies existantes. Cette thèse étudie la famille d’enzymes transglutaminase (TGase) comme biocatalyseur afin d’explorer et d’étendre l’ubiquité et les innovations rendues possibles grâce aux enzymes. Les TGases sont des enzymes versatiles. Leur homologue bactérien, la transglutaminase bactérienne (MTG), est couramment utilisé à l’échelle industrielle pour la transformation alimentaire. Depuis une dizaines d’années, de nombreux efforts ont été faits afin de trouver de nouvelles applications des TGases. En premier lieu, une revue des accomplissements, progrès et défis reliés au développement des TGases sera décrite. Les TGases sont intrinsèquement des catalyseurs de la formation de lien isopeptidiques entre une glutamine et une lysine. Par ce fait, elles ont été initialement testées dans cette thèse pour la synthèse de peptides. Une forme de l’enzyme TGase de mammifères fut en mesure de générer les composés dipeptidiques Gly-Xaa et D-Ala-Gly avec une faible conversion. La MTG possède plusieurs caractéristiques qui font de cette enzyme un candidat intéressant pour le développement de biotechnologies. Elle est stable, non dépendante d’un cofacteur et connait peu de compétition pour sa réaction catalytique inverse. La majeure partie de cette thèse porte exclusivement sur l’utilisation de la MTG. Nous avons développé et caractérisé une réaction chimio-enzymatique en un seul pot pour la conjugaison de peptides et protéines. La présence de glutathion en quantité suffisante permet de contourner l’incompatibilité de la MTG avec le cuivre et ouvre la porte à l’utilisation de la réaction de cycloaddition entre un alcyne et un azoture catalysée par le cuivre, afin d’effectuer le marquage fluorescent de protéines. L’utilisation d’autres méthodes de chimie « click » sans métaux fut aussi étudiée afin d’incorporer divers substrats protéiques. Le marquage de protéines avec la MTG fut investigué de manière combinatoire. Précisément, la ligation de Staudinger, la cycloaddition azoture-alcyne promue par la tension de cycle, ainsi que la ligation de tetrazine (TL) ont été testées. Différents niveaux de conversion ont été atteints, le plus prometteur étant celui obtenu avec la TL. Une étude par cristallographie a été effectuée afin d’élucider comment les substrats contenant une glutamine interagissent avec la MTG. Une méthode de purification alternative de la MTG a été développée afin d’atteindre ce but. Une discussion sur les stratégies et défis est présentée. Finalement, la conjugaison entre un système contenant la MTG comme biocatalyseur de marquage, le domaine B1 de la protéine G (GB1) comme substrat et d’un fluorophore contenant une amine comme sonde fut étudié. Comme deux des constituants de ce système sont des protéines, l’ingénierie d’enzyme peut être entreprise afin d’améliorer leurs propriétés. Une banque de 24 variantes de GB1 fut construite grâce à une approche semi-rationnelle afin d’investiguer quels facteurs sont déterminants pour la sélectivité de la MTG envers la glutamine. Chaque variante étudiée comportait une seule glutamine à une position variable afin d’évaluer l’impact des éléments de structure secondaire où se retrouve la glutamine. L’efficacité pour le marquage a pu être améliorée d’au moins un ordre de grandeur pour huit des substitutions étudiées. Comme chacune des structures secondaires fut marquée, il fut démontré que la MTG n’en préfère pas une en particulier. De plus, la réactivité de la MTG envers la variante I6Q-GB1 fut augmentée en créant des mutations dans son site actif. Ces résultats permettent de comprendre d’avantage la sélectivité de la MTG envers la glutamine, tout en démontrant le potentiel de cette enzyme à être modifiée afin d’être améliorée. / The development of recombinant molecular biology technologies was a turning point for the biological sciences, which has since evolved into dozens upon dozens of different subfields and contributed to extraordinary advances for humans. At the core of many of these advances are the enzymes produced by these techniques, with efforts to understand their form and function laying the groundwork for their application. One of these continuously advancing subfields rooted in enzymology is biocatalysis, in which chemists and engineers embrace biological components and systems to complement, or even replace, existing methodologies. This thesis seeks to further contribute to the advancement and ubiquity of enzymes to be incorporated into future innovations. To this end, transglutaminase (TGase) is the biocatalyst selected for study. TGases are versatile enzymes, with the bacterial homolog, microbial transglutaminase (MTG) being readily used in industrial processes for years, particularly for food processing. An abundance of efforts seeking to apply TGases to other processes have been made within the last decade. We commence by reviewing the accomplishments, progress, and challenges to developing TGase towards new goals. TGase naturally catalyzes the formation of isopeptide bonds utilizing a glutamine and lysine substrates, and one of its first unconventional applications we investigated was for peptide synthesis. We determined the ability and specificity of one form of TGase for various amino acid-derived substrates, observing the formation of Gly-Xaa and D-Ala-Gly dipeptide products, albeit at a low conversion. MTG exhibits several characteristics that make it an appealing candidate for biotechnological development, such as its independence from a cofactor, little competition for its reverse catalytic reaction, and increased stability relative to mammalian TGases. Therefore, the remainder of this thesis pertains exclusively to MTG. We developed and extensively characterized a one-pot chemoenzymatic peptide and protein conjugation scheme. The presence of sufficient glutathione circumvents the incompatibility of the copper-catalyzed azide-alkyne cycloaddition with MTG owing to the presence of copper. We ultimately utilized this chemoenzymatic conjugation scheme for fluorescent protein labeling. We continue to expand upon combinatorial methods to undertake protein labeling by investigating to what extent metal-free click chemistries can be utilized in combination with MTG. Specifically, the Staudinger ligation, strain-promoted azide-alkyne cycloaddition, and tetrazine ligation (TL) were assayed on protein substrates to reveal varying levels of effective conjugation, with the TL being the most promising of the three. The details surrounding the manner in which MTG interacts with its glutamine-containing substrate remains unclear. To address this knowledge gap, we sought to pursue crystallography studies, which required the development a modified purification strategy. We discuss the strategies we investigated and the challenges surrounding such efforts. Finally, we present a conjugation system consisting of MTG as the labeling biocatalyst, the B1 domain of Protein G (GB1) as a substrate, and a small-molecule amine belonging to a recently developed class of fluorophores as a probe. As two components of this system are proteins, enzyme engineering can be applied to further improve their properties. A semi-rational approach was used to generate a 24-member GB1 library to probe the structural determinants of MTG’s glutamine selectivity. Each variant contained a single glutamine at varying positions covering all secondary structure elements, and assayed for reactivity. Eight substitutions resulting in an increased labeling efficiency of at least an order of magnitude were distributed throughout all secondary structure elements, indicating that MTG does not favor one preferentially. In addition, introducing point mutations within MTG’s active site also resulted in increased reactivity towards variant I6Q-GB1. Our results contribute further to understanding the nature of MTG’s glutamine selectivity, while simultaneously demonstrating the potential enzyme engineering has to improve and adjust this system.