Efficient utilization of hydrolysates from steam-exploded gardening residues for lactic acid production by optimization of enzyme addition and pH control.

The expansion of urban green areas has boosted the accumulation of gardening lignocellulosic residues that could be potentially used to produce platform chemicals like lactic acid. However, when using lignocelluloses, pretreatment step, such as steam explosion, is often needed to favour sugar release. Considering that the conversion of glucose from cellulose has been widely addressed, this work is focused on the valorisation of the steam-exploded gardening liquid fraction rich in hemicellulosic sugars. Since oligomeric sugars are usually solubilized during steam explosion, an enzymatic hydrolysis step was required in some cases to increase the monosaccharides content. Although the presence of inhibitors released during pretreatment (e.g. formic acid) hindered hydrolysis yields, the addition of hemicellulases and the enzyme dosage optimization resulted in 85%, 89% and 95% of glucose, xylose and arabinose release from soluble oligomers, respectively. Lactobacillus pentosus CECT4023T was used for lactic acid fermentation of C6 and C5 sugars from the hydrolysate with the highest sugars concentration, that did not require enzymatic hydrolysis. Xylose consumption was hampered due to the inhibitory effect of acids that produced pH drop. Different pH control systems were applied and automatic NaOH addition in bioreactor resulted in 21 g L-1 of lactic acid (95% of the maximum theoretical yield) that implied 44% increase in lactic acid production when compared with flask fermentation. These results provide new insights for the valorisation of emerging lignocellulosic materials like gardening residues into high added-value products.

Assessment of different Bacillus coagulans strains for l-lactic acid production from defined media and gardening hydrolysates: Effect of lignocellulosic inhibitors.

Cellulose valorisation has been successfully addressed for years. However, the use of hemicellulosic hydrolysates is limited due to the presence of C5-sugars and inhibitors formed during pretreatment. Bacillus coagulans is one of the few bacteria able to utilize both C6- and C5-sugars to produce l-lactic acid, but its susceptibility to the lignocellulosic inhibitors needs further investigation. For such a purpose, the tolerance of different B. coagulans strains to increasing concentrations of inhibitors is studied. The isolated A162 strain reached the highest l-lactic acid productivity in all cases (up to 2.4 g L-1  h-1), even in presence of 5 g L-1 of furans and phenols. Remarkably, most of furans and phenolic aldehydes were removed from defined media and hemicellulosic gardening hydrolysate after fermentation with A162. Considering the high productivities and the biodetoxifying effect attained, A162 could be pointed out as a great candidate for valorisation of mixed sugars from hemicellulosic hydrolysates with high inhibitors concentration, promoting the implementation of lignocellulosic biorefineries.

Evolutionary engineering of Lactobacillus pentosus improves lactic acid productivity from xylose-rich media at low pH.

Since xylose is the second most abundant sugar in lignocellulose, using microorganisms able to metabolize it into bio-based chemicals like lactic acid is an attractive approach. In this study, Lactobacillus pentosus CECT4023T was evolved to improve its xylose fermentation capacity even at acid pH by adaptive laboratory evolution in repeated anaerobic batch cultures at increasing xylose concentration. The resulting strain (named MAX2) presented between 1.5 and 2-fold more xylose consumption and lactic acid production than the parental strain in 20 g L-1 xylose defined media independently of the initial pH value. When the pH was controlled in bioreactor, lactic acid productivity at 16 h increased 1.4-fold when MAX2 was grown both in xylose defined media and in wheat straw hydrolysate. These results demonstrated the potential of this new strain to produce lactic acid from hemicellulosic substrates at low pH, reducing the need of using neutralizing agents in the process.

Lactobacillus pentosus CECT 4023 T co-utilizes glucose and xylose to produce lactic acid from wheat straw hydrolysate: Anaerobiosis as a key factor.

Lactic acid is a versatile chemical that can be produced via fermentation of lignocellulosic materials. The heterolactic strain Lactobacillus pentosus CECT 4023 T, that can consume glucose and xylose, was studied to produce lactic acid from steam exploded wheat straw prehydrolysate. The effect of temperature and pH on bacterial growth was analyzed. Besides, the effect of oxygen on lactic acid production was tested and fermentation yields were compared in different scenarios. This strain showed very high tolerance to the inhibitors contained in the wheat straw prehydrolysate. The highest lactic acid yields based on present sugar, around 0.80 g g-1 , were obtained from glucose in presence of 25%, 50%, and 75% v v-1 of prehydrolysate in strict anaerobiosis. Lactic fermentation of wheat straw hydrolysate obtained after enzymatic hydrolysis of the prehydrolysate yielded 0.39 g of lactic acid per gram of released sugars, which demonstrated the high potential of L. pentosus to produce lactic acid from hemicellulosic hydrolysates. Results presented herein not only corroborated the ability of L. pentosus to grow using mixtures of sugars, but also demonstrated the suitability of this strain to be applied as an efficient lactic acid producer in a lignocellulosic biorefinery approach. © 2018 American Institute of Chemical Engineers Biotechnol. Prog., 35: e2739, 2019.