UCL-3500紫外交聯(lián)儀在紫外線誘導(dǎo)突變中的應(yīng)用

摘要：

2025 年，北京化工大學(xué)化學(xué)工程學(xué)院聯(lián)合生命科學(xué)與技術(shù)學(xué)院綠色生物制造國家重點實驗室在《Biochemical Engineering Journal》期刊發(fā)表文獻(xiàn)《Screening of high-yielding β-farnesene Saccharomyces cerevisiae strains and quantification of fermentation processes: A new fluorescence-based high-throughput screening and quantification platform》，該文獻(xiàn)明確指出利用紫外交聯(lián)儀 UCL-3500 可高效開展紫外線誘導(dǎo)突變實驗，為高產(chǎn) β- 法尼烯釀酒酵母菌株的篩選及發(fā)酵過程量化提供了關(guān)鍵技術(shù)支撐，顯著提升了紫外線誘導(dǎo)突變篩選的效率與準(zhǔn)確性。

紫外交聯(lián)儀 UCL-3500 助力紫外線誘導(dǎo)突變篩選高產(chǎn)酵母菌株

2025 年，北京化工大學(xué)主導(dǎo)的研究團(tuán)隊在《Biochemical Engineering Journal》發(fā)表重要成果，成功建立了基于熒光探針的高通量篩選與定量平臺，實現(xiàn)了從紫外線誘導(dǎo)突變菌株篩選到規(guī)模化發(fā)酵過程監(jiān)測的全流程優(yōu)化。在該研究中，紫外交聯(lián)儀 UCL-3500 作為核心實驗設(shè)備，承擔(dān)了釀酒酵母的紫外線誘導(dǎo)突變?nèi)蝿?wù)，其穩(wěn)定的紫外線輸出、精準(zhǔn)的照射時間控制能力，為構(gòu)建高質(zhì)量突變體庫、高效篩選高產(chǎn) β- 法尼烯菌株奠定了堅實基礎(chǔ)。

北京化工大學(xué)化學(xué)工程學(xué)院與綠色生物制造國家重點實驗室是我國生物化工領(lǐng)域的重要研究陣地，圍繞綠色可持續(xù)生產(chǎn)的重大戰(zhàn)略需求，在微生物代謝工程、生物催化與轉(zhuǎn)化、發(fā)酵過程優(yōu)化等前沿領(lǐng)域開展系統(tǒng)研究。重點方向包括萜類化合物生物合成、微生物菌株改良、高效檢測技術(shù)開發(fā)等，曾在多個國際優(yōu)秀期刊發(fā)表多項突破性成果，為生物制造產(chǎn)業(yè)的升級發(fā)展提供了重要理論支撐和技術(shù)儲備。

在該實驗中，研究團(tuán)隊以產(chǎn) β- 法尼烯工程釀酒酵母菌株 BUHY0003 為研究對象，采用紫外交聯(lián)儀 UCL-3500 開展紫外線誘導(dǎo)突變實驗。研究人員將 5mL 菌株懸液置于無菌培養(yǎng)皿中，加入無菌磁力攪拌子使菌液形成均勻薄層，隨后將培養(yǎng)皿置于紫外交聯(lián)儀 UCL-3500 紫外線燈下方約 20cm 處，設(shè)置 0、20、25、30 等多個照射時間梯度進(jìn)行紫外線誘導(dǎo)突變處理，每個處理組均設(shè)置三次重復(fù)。紫外線誘導(dǎo)突變處理后，將菌液在黑暗條件下離心收集，重懸于 YPD 培養(yǎng)基中恢復(fù)培養(yǎng) 2 小時，再涂布于 YPD 瓊脂培養(yǎng)基上培養(yǎng)，通過計算菌落存活率確定紫外線誘導(dǎo)突變參數(shù)。最終發(fā)現(xiàn)，當(dāng)紫外交聯(lián)儀 UCL-3500 的照射時間為 50 秒時，菌株致死率達(dá)到 94.76%，此條件下既能保證足夠的突變負(fù)荷，又能維持突變體庫的實用規(guī)模，是開展紫外線誘導(dǎo)突變的設(shè)置。

通過紫外交聯(lián)儀 UCL-3500 完成紫外線誘導(dǎo)突變后，研究團(tuán)隊利用構(gòu)建的熒光高通量篩選平臺對突變菌株進(jìn)行篩選，成功獲得高產(chǎn)突變株 BUHY0004，其 β- 法尼烯產(chǎn)量較原始菌株提升 37.34%。后續(xù)經(jīng) 5L 和 30L 補(bǔ)料分批發(fā)酵驗證，該突變株表現(xiàn)出良好的規(guī)模化生產(chǎn)性能，產(chǎn)量分別達(dá)到 35.1g/L 和 33.3g/L，充分證明了紫外交聯(lián)儀 UCL-3500 在紫外線誘導(dǎo)突變菌株改良中的可靠性與高效性。

紫外交聯(lián)儀 UCL-3500 是一款專為微生物紫外線誘導(dǎo)突變設(shè)計的專業(yè)設(shè)備，采用穩(wěn)定的紫外線光源設(shè)計，可提供均勻、高強(qiáng)度的紫外線照射，確保紫外線誘導(dǎo)突變的有效性。設(shè)備支持靈活的照射時間調(diào)節(jié)與能量調(diào)控，能滿足不同菌株的紫外線誘導(dǎo)突變需求，可精準(zhǔn)控制突變強(qiáng)度，為篩選有益突變體創(chuàng)造有利條件。操作簡便，無需復(fù)雜的前期調(diào)試，適用于實驗室大規(guī)模突變體庫構(gòu)建。其適配性強(qiáng)，可廣泛應(yīng)用于釀酒酵母、大腸桿菌等多種微生物的紫外線誘導(dǎo)突變實驗，為代謝工程、菌株改良、發(fā)酵工業(yè)等領(lǐng)域的研究提供強(qiáng)有力的技術(shù)支持。目前，該設(shè)備已成為高校、科研院所開展微生物紫外線誘導(dǎo)突變相關(guān)研究的重要工具，相關(guān)研究成果已在多個國際優(yōu)秀期刊發(fā)表。

原文相關(guān)段落摘錄：

Saccharomyces cerevisiae is regarded as one of the most important model strain and industrial workhorses in metabolic engineering and biotechnology, and has been extensively employed for the manufacturing of natural products. Although substantial advances in genome editing and rational design have been achieved, growth retardation in scale-up fermentations is frequently observed in engineered yeasts owing to the complex regulation of central carbon metabolism. In this context, ultraviolet (UV) mutagenesis is recognized as a powerful complementary strategy, enabling the rapid generation of extensive mutant libraries and facilitating the identification of beneficial alleles.

UV mutagenesis and 96-well plate screeningIn this study, the engineered S. cerevisiae strain BUHY0003 used for producing β-farnesene was constructed according to our previous work. A mutagenesis chamber (UCL-3500, Shanghai Luyang Biotechnology Co., Ltd., China) equipped with two 18 W UV lamps was used as the light source and was preheated for 20 min before use. 5 mL of BUHY0003 strain suspension were placed in a sterile 9 cm culture dish. A sterile 1.5 cm magnetic stir bar was added, and the dish was gently agitated on a shaker to form a uniform thin layer. The dish was positioned approximately 20 cm below the UV lamps and was irradiated at normal incidence in triplicate. Irradiation times were set to 0, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, and 80 s. Following irradiation, cells were collected by centrifugation at 4000 × g for 4 min in the dark. The supernatant was discarded, and the cells were resuspended in YPD medium followed by a 2-hour recovery incubation at 30?C with shaking at 200 rpm. 100 μL of the suspension were spread onto YPD agar medium and were incubated inverted at 30?C for 96 h. Under sterile conditions, colonies were counted and lethality was calculated.

DOI：10.1016/j.bej.2025.110006