Comparative proteomic analysis of Rhizopus oryzae hyphae displaying filamentous and pellet morphology
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ORIGINAL ARTICLE
Comparative proteomic analysis of Rhizopus oryzae hyphae displaying filamentous and pellet morphology Longfei Yin1 · Xi Luo1 · Yingying Zhang1 · Weilong Zheng1 · Fengwei Yin1 · Yongqian Fu1 Received: 29 April 2020 / Accepted: 28 September 2020 © King Abdulaziz City for Science and Technology 2020
Abstract Industrial strains of Rhizopus oryzae is known for its strong ability to produce L-( +)-lactic acid, ethanol, and fumaric acid at high yields. To better understand the underlying mechanism behind the physiology of R. oryzae, we conducted the proteome changes between two different morphologies using two-dimensional polyacrylamide gel electrophoresis and mass spectrometry. R. oryzae exhibited pellet morphology and filamentous morphology when the initial pH of the culture medium was 3.0 and 5.0, respectively. The concentration of lactic acid reached 63.5 g L−1 in the samples containing the pellet morphology, compared to 41.5 g L −1 produced by filamentous R. oryzae. Proteomic analysis indicated that expression levels of 128 proteins changed significantly. Of these, 17 protein spots were successfully identified by mass spectrometry and were deemed to be mainly involved in carbohydrate metabolism, genetic information processing, chitin metabolism, protein catabolism, protein folding, and antioxidative pathway. L-lactate dehydrogenase (RO3G_06188), enolase (RO3G_05466) and 2, 3-bisphosphoglycerate-independent phosphoglycerate mutase (RO3G_02462) were found to be upregulated, while isocitrate dehydrogenase (RO3G_13820) was downregulated in the samples with pellet morphology compared to the filamentous hyphae. These results suggested that more carbon flow was directed towards lactic acid biosynthesis in R. oryzae hyphae with pellet morphology. Keywords Rhizopus oryzae · Morphology · Proteomics · Lactic acid
Introduction The morphology of filamentous fungi has received attention in terms of both pure and applied science. The filamentous mycelia of Rhizopus species can grow in different morphological forms such as clumps, filaments, and pellets, depending on the growth conditions (Kossen 2000). The particular morphology is determined not only by the genetic makeup of the fungal species but also the nature of inoculum as well as the chemical (medium constituents) and physical (pH, temperature, mechanical forces) culture conditions (Kossen 2000). Moreover, the morphology of filamentous fungi during fermentation is critical for maximum performance. Although the optimal morphology for the production of various chemicals differs among fungi, pellets are often the preferable morphological forms in industrial * Yongqian Fu [email protected] 1
Institute of Biomass Resources, Taizhou University, Taizhou 318000, China
fermentation processes. For example, pellet morphology is necessary for maximum production of fumaric acid by R. oryzae 1526 (Das and Brar 2014), L-( +)-lactic acid by R. oryzae NRRL 395 (Liu et al. 2006) and heterologous proteins by Aspergillus niger (Xu et al. 2000). Our study also show
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