Novel differential scanning calorimetry (DSC) application to select polyhydroxyalkanoate (PHA) producers correlating 3-hydroxyhexanoate (3-HHx) monomer with melting enthalpy.

IF 3.5 3区生物学 Q2 BIOTECHNOLOGY & APPLIED MICROBIOLOGY Bioprocess and Biosystems Engineering Pub Date : 2024-10-01 Epub Date: 2024-08-06 DOI:10.1007/s00449-024-03054-9

Hee Ju Jung, Byungchan Kim, Tae-Rim Choi, Suk Jin Oh, Suwon Kim, Yeda Lee, Yuni Shin, Suhye Choi, Jinok Oh, So Yeon Park, Young Sik Lee, Young Heon Choi, Yung-Hun Yang

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Abstract

Polyhydroxyalkanoate (PHA) is an environmental alternative to petroleum-based plastics because of its biodegradability. The polymer properties of PHA have been improved by the incorporation of different monomers. Traditionally, the monomer composition of PHA has been analyzed using gas chromatography (GC) and nuclear magnetic resonance (NMR), providing accurate monomer composition. However, sequential analysis of the thermal properties of PHA using differential scanning calorimetry (DSC) remains necessary, providing crucial insights into its thermal characteristics. To shorten the monomer composition and thermal property analysis, we directly applied DSC to the analysis of the obtained PHA film and observed a high correlation (r² = 0.98) between melting enthalpy and the 3-hydroxyhexanoate (3-HHx) mole fraction in the polymer. A higher 3-HHx fraction resulted in a lower melting enthalpy as 3-HHx provided the polymer with higher flexibility. Based on this, we selected the poly(3-hydroxybutyrate-co-3-hydroxyhexanoate) (P(3HB-co-3HHx)) producing strain from Cupriavidus strains that newly screened and transformed with vectors containing P(3HB-co-3HHx) biosynthetic genes, achieving an average error rate below 1.8% between GC and DSC results. Cupriavidus sp. BK2 showed a high 3-HHx mole fraction, up to 10.38 mol%, with T_m(℃) = 171.5 and ΔH of T_m (J/g) = 48.0, simultaneously detected via DSC. This study is an example of the expansion of DSC for PHA analysis from polymer science to microbial engineering.

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应用新型差示扫描量热法 (DSC) 挑选聚羟基烷酸酯 (PHA) 生产商，将 3-hydroxyhexanoate (3-HHx) 单体与熔化焓相关联。

聚羟基烷酸酯（PHA）具有生物降解性，是石油基塑料的环保替代品。通过加入不同的单体，PHA 的聚合物特性得到了改善。传统上，PHA 的单体组成是通过气相色谱法（GC）和核磁共振法（NMR）进行分析，从而提供准确的单体组成。然而，使用差示扫描量热仪（DSC）对 PHA 的热特性进行连续分析仍然是必要的，这将为深入了解其热特性提供重要依据。为了缩短单体成分和热特性分析的时间，我们直接使用 DSC 分析所获得的 PHA 薄膜，并观察到熔化焓与聚合物中 3-hydroxyhexanoate (3-HHx) 分子分数之间存在高度相关性（r2 = 0.98）。3-HHx 分数越高，熔化焓越低，因为 3-HHx 使聚合物具有更高的柔韧性。基于这一点，我们从新筛选并用含有 P（3HB-co-3HHx）生物合成基因的载体转化的铜绿微囊藻菌株中选出了生产聚（3-羟基丁酸-co-3-羟基己酸）（P（3HB-co-3HHx））的菌株，使 GC 和 DSC 结果之间的平均误差率低于 1.8%。Cupriavidus sp. BK2 表现出较高的 3-HHx 分子分数，高达 10.38 摩尔%，Tm (℃) = 171.5，Tm 的 ΔH (J/g) = 48.0，同时通过 DSC 检测到。这项研究是将 DSC 用于 PHA 分析从聚合物科学扩展到微生物工程的一个范例。

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来源期刊

Bioprocess and Biosystems Engineering 工程技术-工程：化工

CiteScore

7.90

自引率

2.60%

发文量

147

审稿时长

2.6 months

期刊介绍： Bioprocess and Biosystems Engineering provides an international peer-reviewed forum to facilitate the discussion between engineering and biological science to find efficient solutions in the development and improvement of bioprocesses. The aim of the journal is to focus more attention on the multidisciplinary approaches for integrative bioprocess design. Of special interest are the rational manipulation of biosystems through metabolic engineering techniques to provide new biocatalysts as well as the model based design of bioprocesses (up-stream processing, bioreactor operation and downstream processing) that will lead to new and sustainable production processes. Contributions are targeted at new approaches for rational and evolutive design of cellular systems by taking into account the environment and constraints of technical production processes, integration of recombinant technology and process design, as well as new hybrid intersections such as bioinformatics and process systems engineering. Manuscripts concerning the design, simulation, experimental validation, control, and economic as well as ecological evaluation of novel processes using biosystems or parts thereof (e.g., enzymes, microorganisms, mammalian cells, plant cells, or tissue), their related products, or technical devices are also encouraged. The Editors will consider papers for publication based on novelty, their impact on biotechnological production and their contribution to the advancement of bioprocess and biosystems engineering science. Submission of papers dealing with routine aspects of bioprocess engineering (e.g., routine application of established methodologies, and description of established equipment) are discouraged.