SBIR Phase I: A Novel Probe for Biofilms

SBIR 第一阶段：生物膜的新型探针

• 批准号: 2110505
• 负责人: Nancy Kravit
• 金额: $ 25.59万
• 依托单位: Tethys Research LLC
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-08-15 至 2023-07-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=2110505&HistoricalAwards=false
• 关键词: SBIR; Phase; Novel; Probe; Biofilms

The broader impact of this Small Business Innovation Research (SBIR) Phase I project will be to improve performance of pulp and paper mills, especially small mills in rural areas, and to improve the quality of their products. The main components of wood are cellulose (used for paper), hemicellulose, and lignin (which makes paper turn yellow and brittle with age). Removal of lignin is key to papermaking, but it is typically attached to the hemicellulose. The project advances a technology to measure how much lignin is left in wood pulp. In the longer term, the probe may also detect biofilms which are layers of carbohydrates and other materials in which bacteria and fungi can grow. Water treatment facilities and processors of plant-derived materials routinely find that microorganisms naturally present in their starting materials form biofilms that can accumulate on machinery or contaminate the products. This technology has wide-ranging applications. The proposed project creates a peptide probe for non-glycosidic bonds to xylose residues of oligo- or polysaccharides. These bonds are known to exist in wood, and may exist in other woody plants and biofilms. The project employs the only enzyme known to break these bonds, XLE (xylan:lignin etherase) in order to create a peptide probe to indicate their presence. XLE’s homology to known xylosidases has permitted the identification of residues very likely to participate in the catalytic activity. These residues will be targets of modification that remove catalytic activity while retaining the ability to bind to XLE’s non-glycosidic substrate. The modifications will be performed using standard techniques of molecular biology and evaluated using with a novel indicating substrate for XLE activity. XLE’s homology with known glycosidases has also allowed identification of the enzyme’s potential binding site for its substrate. A peptide with an attached visible marker corresponding to this sequence but including the sequence changes that remove the catalytic activity will be synthesized. The resulting peptide will be tested to see if it reacts with wood pulp and biofilms.This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.

这一小企业创新研究（SBIR）第一阶段项目的更广泛影响将是提高纸浆和造纸米尔斯厂，特别是农村地区的小型米尔斯厂的业绩，并提高其产品质量。 木材的主要成分是纤维素（用于造纸），半纤维素和木质素（使纸张随着时间变黄变脆）。 去除木质素是造纸的关键，但它通常附着在半纤维素上。 该项目推进了一项技术，以测量多少木质素留在木浆。 从长远来看，探针还可以检测生物膜，生物膜是碳水化合物层和细菌和真菌可以生长的其他材料。 水处理设施和植物源材料的加工者通常会发现，在其起始材料中天然存在的微生物会形成生物膜，这些生物膜可能会积聚在机器上或污染产品。这项技术有着广泛的应用。 拟议的项目创建一个肽探针的非糖苷键的木糖残基的寡糖或多糖。 已知这些键存在于木材中，并且可能存在于其他木本植物和生物膜中。 该项目采用了唯一已知的酶来打破这些键，XLE（木聚糖：木质素醚酶），以创建一个肽探针来指示它们的存在。 XLE与已知的木糖醇酶的同源性已经允许鉴定非常可能参与催化活性的残基。 这些残基将是修饰的靶标，其去除催化活性，同时保留与XLE的非糖苷底物结合的能力。 将使用分子生物学的标准技术进行修饰，并使用XLE活性的新型指示底物进行评价。 XLE与已知的糖苷酶的同源性也允许鉴定酶对其底物的潜在结合位点。 将合成具有对应于该序列但包括去除催化活性的序列变化的连接的可见标记物的肽。 该奖项反映了NSF的法定使命，并通过使用基金会的知识价值和更广泛的影响审查标准进行评估，被认为值得支持。