Novel extremophilic enzymes for new applications in Healthcare Products (Ref: 4188)

对细胞通讯和微生物生物膜形成的了解是联合利华对其许多家用和保健产品的关键兴趣，这些产品希望针对这一领域来改善卫生。酶可以通过特定的微生物通讯内酯分子的水解来干扰生物膜的形成，这对解决这些问题是关键的。将这种方法与其他降解细胞壁的水解酶和清洁化学物质相结合的潜力，试图创建一种更全面的方法，以提供所需的结果。如果多重干预模式是成功的，它也可能有助于预防不受欢迎的细菌耐药性。这与联合利华的清洁未来议程和提高产品的可持续性很好地结合在一起。耐热内酯酶已经使用来自高温环境的结构生物信息学和来自炎热环境的超基因组鉴定。这三类乳糖酶具有不同的结构和不同的酶机制。使用这些酶来分解污染的生物膜比使用抗生素有优势，并且重要的是消除了抗生素耐药性的问题。群体感应内切酶已被报道破坏生物膜的形成，烯醇内酯酶是破坏生物膜的新的潜在靶点，关于这些内切酶的底物专一性的一些初步信息是可用的。据报道，葡萄糖内酯酶还可以破坏生物膜的形成。最近的研究发现，许多酶对改善保健产品有用。传统上，迄今为止使用的大多数工业酶都是从中温生物获得的，这些生物在恶劣的工业条件下稳定性有限，包括高温、高盐浓度、极端pH和配方化学品，如表面活性剂。因此，发现强健的酶，设计更活跃的变体，以及更好地了解它们的分子机制，是未来工业生物技术中生物催化策略发展的一些关键挑战。极端微生物是工业生物催化剂的一个有吸引力的来源，因为它们是自然进化出来的强大的酶，能够在极端条件下发挥作用。此外，在一种环境(如高温)中发现的极端嗜热酶通常也能耐受其他极端条件(如有机溶剂、表面活性剂和其他化学制剂成分)，使其可用于各种应用。任务1--使用群体感应乳糖酶、烯醇乳糖酶、葡萄糖内酯酶和AHL酰基酶在微生物生物膜的分解中评估新的乳糖酶和酰基酶。任务2--在Metagenome DNA数据库中搜索进一步相关的酶。通过X射线结晶学和分子建模/底物对接对选定的酶进行结构表征。晶体结构将被用来了解这些酶的底物选择性和催化机理，并指导蛋白质工程实验，以提高它们在工业条件下的效率。任务3-评估选定的酶应用于生物膜破坏。在材料创新工厂内的联合利华研发实验室的项目布置将涉及使用定制的自动化微生物测试平台(针对表面附着的微生物)，以及使用射流设备和可视化技术来观察和测量生物膜的发展，包括基因报告系统。任务4-所有对生物膜的形成或破坏具有积极影响的潜在内酰胺酶将受到不同类型的固定化技术的影响，以帮助开发和扩大其在家庭和保健产品中的潜在应用。

The understanding of cellular communication and microbial biofilm formation is a key interest for Unilever with respect to many of their home and healthcare products which look to target this area for improving hygiene. Enzymes that can interfere with the biofilm formation by specifically hydrolysing the microbial communication lactone molecules are of key importance towards addressing these problems. The potential of combining this approach with other cell wall degrading hydrolases and cleaning chemistries, attempts to create a more holistic approach to multiple modes of action providing the desired outcome. If the multiple mode of intervention is successful it may also help in the prevention of undesired bacterial resistance. This is well aligned to the Clean Future agenda of Unilever and increasing sustainability of products.Thermostable lactonases have been identified using structural bioinformatics from thermophilic and metagenomes from 'Hot Environments'. The three classes of lactonase enzymes have different structures and different enzymatic mechanisms. The use of these enzymes to break down contaminating biofilms has advantages over the use of antibiotics, and importantly eliminates problems with antibiotic resistance. The Quorum sensing lactonases have been reported to disrupt biofilm formation, the enol lactonases are a new potential target for biofilm disruption and some preliminary information on substrate specificity of these lactonases is available. The Gluconolactonases have been reported to also disrupt biofilm formation.Recent studies have identified many enzymes useful for improved Healthcare products. Traditionally the majority of industrial enzymes used to date have been obtained from mesophilic organisms, which have a limited stability under harsh industrial conditions including high temperatures, high salt concentrations, extreme pH, and formulation chemicals such as surfactants. Therefore, the discovery of robust enzymes, engineering of more active variants, as well as better understanding of their molecular mechanisms, represent some of the key challenges for the development of future biocatalytic strategies in Industrial Biotechnology. The extremophilic microorganisms represent an attractive source of industrial biocatalysts because they are naturally evolved robust enzymes to function under extreme conditions. In addition, extremophilic enzymes found in one environment (e.g. high temperature) are typically also tolerant to other extreme conditions (e.g. organic solvents, surfactants and other chemical formulation components) making them useful for a variety of applications.Task 1 - Assessment of novel lactonase and acylases in breakdown of microbial biofilms using quorum sensing lactonases, enol lactonases, gluconolactonases and AHL acylases.Task 2 - Search Metagenomics DNA databases for further related enzymes. Carry out structurally characterization of selected enzymes by both X-ray crystallography and molecular modelling/ substrate docking. Crystal structures will be used to understand the substrate selectivity and catalytic mechanism of these enzymes and to direct protein engineering experiments to improve their efficacy under industrial conditions.Task 3 - Evaluation of selected enzyme for application in biofilm disruption. Project placements at Unilever R&D labs within the Material Innovation Factory will involve the use of a bespoke automated microbiological testing platform (for surface adhered microbes), as well as use of fluidic devices and visualisation techniques to observe and measure biofilm development, including gene reporter systems.Task 4 - All potential lactonases that have positive effects on the formation or disruption of biofilms will be subjected to different types of immobilisation techniques to help develop and broaden their potential applications for home and healthcare products.