Pomraning Kyle R, Collett James R, Kim Joonhoon, Panisko Ellen A, Culley David E, Dai Ziyu, Deng Shuang, Hofstad Beth A, Butcher Mark G, Magnuson Jon K
1Pacific Northwest National Laboratory, Richland, WA USA.
2Joint BioEnergy Institute, Emeryville, CA USA.
Biotechnol Biofuels. 2019 Jun 26;12:162. doi: 10.1186/s13068-019-1510-z. eCollection 2019.
Efficient and economically viable production of biofuels from lignocellulosic biomass is dependent on mechanical and chemical pretreatment and enzymatic hydrolysis of plant material. These processing steps yield simple sugars as well as plant-derived and process-added organic acids, sugar-derived dehydration products, aldehydes, phenolics and other compounds that inhibit the growth of many microorganisms. is an oleaginous yeast capable of robust growth on a variety of sugars and lipid accumulation on pretreated lignocellulosic substrates making it attractive as an industrial producer of biofuels. Here, we examined gene expression during batch growth and lipid accumulation in a 20-L bioreactor with either a blend of pure glucose and xylose or pretreated corn stover (PCS) that had been enzymatically hydrolyzed as the carbon sources.
We monitored sugar and ammonium utilization as well as biomass accumulation and found that growth of is inhibited with PCS hydrolysate as the carbon source. Both acetic acid and furfural are present at concentrations toxic to in PCS hydrolysate. We quantified gene expression at seven time-points for each carbon source during batch growth and found that gene expression is similar at physiologically equivalent points. Analysis of promoter regions revealed that gene expression during the transition to lipid accumulation is regulated by carbon and nitrogen catabolite repression, regardless of carbon source and is associated with decreased expression of the translation machinery and suppression of the cell cycle. We identified 73 differentially expressed genes during growth phase in the bioreactor that may be involved in detoxification of corn stover hydrolysate.
Growth of is inhibited by compounds present in PCS hydrolysate. Here, we monitored key metabolites to establish physiologically equivalent comparisons during a batch bioreactor run comparing PCS hydrolysate and purified sugars. 's response to PCS hydrolysate is primarily at the beginning of the run during growth phase when inhibitory compounds are presumably at their highest concentration and inducing the general detoxification response by . Differentially expressed genes identified herein during growth phase will aid in the improvement of industrial strains capable of robust growth on substrates containing various growth inhibitory compounds.
从木质纤维素生物质高效且经济可行地生产生物燃料取决于植物材料的机械和化学预处理以及酶水解。这些加工步骤产生单糖以及植物衍生和加工过程中添加的有机酸、糖衍生的脱水产物、醛、酚类和其他抑制许多微生物生长的化合物。解脂耶氏酵母是一种产油酵母,能够在多种糖类上旺盛生长,并在预处理的木质纤维素底物上积累脂质,使其成为有吸引力的生物燃料工业生产菌株。在此,我们研究了在20升生物反应器中分批生长和脂质积累过程中的基因表达情况,该反应器以纯葡萄糖和木糖的混合物或经酶水解的预处理玉米秸秆(PCS)作为碳源。
我们监测了糖和铵的利用以及生物量积累,发现以PCS水解产物作为碳源时,解脂耶氏酵母的生长受到抑制。乙酸和糠醛在PCS水解产物中的浓度对解脂耶氏酵母有毒。我们在分批生长过程中对每种碳源的七个时间点的基因表达进行了定量,发现基因表达在生理等效点相似。启动子区域分析表明,无论碳源如何,向脂质积累转变过程中的基因表达受碳和氮分解代谢物阻遏调控,并且与翻译机制表达降低和细胞周期抑制相关。我们鉴定出生物反应器生长阶段有73个差异表达基因,这些基因可能参与玉米秸秆水解产物的解毒。
解脂耶氏酵母的生长受到PCS水解产物中存在的化合物的抑制。在此,我们监测了关键代谢物,以便在分批生物反应器运行过程中建立PCS水解产物和纯化糖之间的生理等效比较。解脂耶氏酵母对PCS水解产物的反应主要在运行开始的生长阶段,此时抑制性化合物的浓度可能最高,并诱导解脂耶氏酵母产生一般解毒反应。本文在生长阶段鉴定出的差异表达基因将有助于改进能够在含有各种生长抑制化合物的底物上旺盛生长的工业菌株。