Current Progress in Production of Building-Block Organic Acids by Consolidated Bioprocessing of Lignocellulose

Several organic acids have been indicated among the top value chemicals from biomass. Lignocellulose is among the most attractive feedstocks for biorefining processes owing to its high abundance and low cost. However, its highly complex nature and recalcitrance to biodegradation hinder development of cost-competitive fermentation processes. Here, current progress in development of single-pot fermentation (i.e., consolidated bioprocessing, CBP) of lignocellulosic biomass to high value organic acids will be examined, based on the potential of this approach to dramatically reduce process costs. Different strategies for CBP development will be considered such as: (i) design of microbial consortia consisting of (hemi)cellulolytic and valuable-compound producing strains; (ii) engineering of microorganisms that combine biomass-degrading and high-value compound-producing properties in a single strain. The present review will mainly focus on production of organic acids with application as building block chemicals (e.g., adipic, cis,cis-muconic, fumaric, itaconic, lactic, malic, and succinic acid) since polymer synthesis constitutes the largest sector in the chemical industry. Current research advances will be illustrated together with challenges and perspectives for future investigations. In addition, attention will be dedicated to development of acid tolerant microorganisms, an essential feature for improving titer and productivity of fermentative production of acids.

Metabolic engineering of methylotrophic Pichia pastoris for the production of β-alanine

Abstractβ-Alanine (3-aminopropionic acid) is the only naturally occurring β-amino acid and an important precursor for the synthesis of a variety of nitrogen-containing chemicals. Fermentative production of β-alanine from renewable feedstocks such as glucose has attracted significant interest in recent years. Methanol has become an emerging and promising renewable feedstock for biomanufacturing as an alternative to glucose. In this work, we demonstrated the feasibility of β-alanine production from methanol using Pichia pastoris (Komagataella phaffii) as a methylotrophic cell factory. L-Aspartate-α-decarboxylases (ADCs) from different sources were screened and expressed in P. pastoris, followed by the optimization of aspartate decarboxylation by increasing the ADC copy number and C4 precursor supply via the overexpression of aspartate dehydrogenase. The production potential of the best strain was further evaluated in a 1-L fermenter, and a β-alanine titer of 5.6 g/L was obtained. To our best knowledge, this is the highest metabolite production titer ever reached in P. pastoris using methanol as the substrate. Graphic abstract

Fermentative Production of Lasiodiplodan by Lasiodiplodia theobromae CCT3966 from Pretreated Sugarcane Straw

Lasiodiplodan is a β-glucan polymer with different interesting characteristics, including therapeutic properties. It is an extracellular product, which is produced by the filamentous fungus Lasiodiplodia theobromae, using glucose as a substrate. In the present work, the production of lasiodiplodan was studied by the utilization of sugarcane straw as a low-cost carbon source. Glucose-rich sugarcane straw hydrolysate was obtained by a sequential pretreatment with dilute nitric acid (1% v/v) and sodium hydroxide (1% w/v), followed by enzymatic hydrolysis. The fermentation process was conducted by the cultivation of the strain Lasiodiplodia theobromae CCT3966 in sugarcane straw hydrolysate in a shake flask at 28 °C for 114 h. It was found that hydrolysate obtained after enzymatic hydrolysis contained 47.10 gL−1 of glucose. Fermentation experiments of lasiodiplodan synthesis showed that the peak yield and productivity of 0.054 gg−1 glucose consumed and 0.016 gL−1 h−1, respectively, were obtained at 72 h fermentation time. Fungal growth, glucose consumption, and lasiodiplodan production from sugarcane straw hydrolysate presented a similar pattern to kinetic models. The study on the chemical structure of lasiodiplodan produced showed it had a β-glucan construction. The current study revealed that sugarcane straw is a promising substrate for the production of lasiodiplodan.

Biorefinery Concept Employing Bacillus coagulans: LX-Lignin and L-(+)-Lactic Acid from Lignocellulose

A new biorefinery concept is proposed that integrates the novel LX-Pretreatment with the fermentative production of L-(+)-lactic acid. Lignocellulose was chosen as a substrate that does not compete with the provision of food or feed. Furthermore, it contains lignin, a promising new chemical building material which is the largest renewable source for aromatic compounds. Two substrates were investigated: rye straw (RS) as a residue from agriculture, as well as the fibrous digestate of an anaerobic biogas plant operated with energy corn (DCS). Besides the prior production of biogas from energy corn, chemically exploitable LX-Lignin was produced from both sources, creating a product with a low carbohydrate and ash content (90.3% and 88.2% of acid insoluble lignin). Regarding the cellulose fraction of the biomass, enzymatic hydrolysis and fermentation experiments were conducted, comparing a separate (SHF), simultaneous (SSF) and prehydrolyzed simultaneous saccharification and fermentation (PSSF) approach. For this purpose, thermophilic B. coagulans 14-300 was utilized, reaching 38.0 g L−1 LA in 32 h SSF from pretreated RS and 18.3 g L−1 LA in 30 h PSSF from pretreated DCS with optical purities of 99%.

Microbial lipid fermentation of Trichosporon cutaneum in high saline water

Fermentative production of microbial lipid requires high fresh water input. The utilization of high saline seawater or industrial wastewater is an important alternative to reduce the freshwater consumption. This study revealed that oleaginous yeast Trichosporon cutaneum was tolerant to a high salinity up to 130 g/L of NaCl after long-term adaptive evolution. Lipid fermentation of T. cutaneum in seawater achieved the lipid production of 31.7 g/L with approximately 36% greater than that in freshwater. The saline water containing phenol was also tested for lipid fermentation and 23.6 g/L of lipid was produced simultaneously with the complete biodegradation of phenol. An interesting phenomenon was also observed that the yeast cells spontaneously segregated onto the upper surface of the saline water. This study extended the lipid fermentation options with practical application potentials.