Continuous fermentation of clarified corn stover hydrolysate for the production of lactic acid at high yield and productivity

The present study investigated the synergistic effect of nutritional supplements (amino acid and Tween 80) on lactic acid production by Lactobacillus delbruckii utilizing a sugar refinery by product (cane molasses) in a submerged fermentation process. Initially, the effect of individual factors on lactic acid yield was studied by supplementing amino acids and their combinations, Tween 80 and cane molasses at varying concentrations in production medium. A combination of l-phenylalanine and l-lysine gave a maximum lactic acid yield of 47.89 ± 0.1 g/L on a dry cell weight basis at individual factor level. Similarly, maximum lactic acid yield was obtained by supplementing the production medium with 40.0 g/L and 2.0 g/L Tween 80 and cane molasses, respectively, at individual factor level. In order to further improve the lactic acid yield, nutritional supplements were optimized by central composite rotatable design (CCRD) using Minitab 15 software. Shake flask cultivation under optimized conditions, i.e., cane molasses (32.40 g/L), Tween 80 (2.0 g/L) and l-phenylalanine and l-lysine (34.0 mg/L) gave a lactic acid yield of 64.86 ± 0.2 g/L, corresponding to 95.0 % of the predicted yield of 67.78 ± 0.3 g/L. Batch cultivation performed in 7.5 L bioreactor (working volume: 3.0 L) under optimized conditions gave maximum lactic acid yield and productivity of 79.12 ± 0.2 g/L and 3.40 g/L·h, which is higher than previous studies with reduced fermentation time. Screening of lactic acid producing bacteria and characterization of lactic acid was also done.

The continuous production of lactic acid from deproteinized whey by immobilized single and mixed culture of L. casei and L. lactis in Ca‐alginate beads has been investigated. A coimmobilized culture system gave better results than immobilized single cultures regarding lactic acid concentration, productivity, yield, and lactose utilization. Maximum lactic acid productivity of 7 g/lh was obtained at D=0.4 h−1 with a yield of 70% lactic acid and 50% lactose utilization. At a dilution rate of 0.1 h−1, a lactic acid productivity of 2.5 g/lh was obtained with a 55.5% lactic acid yield and 90% lactose utilization. The bioreactor system was operated at a constant dilution rate of 0.1 h−1 for 20 days without loss of original activity. In this case, the average lactic acid productivity, lactic acid yield and lactose utilization were 24 g/lh, 55% and 90%, respectively.

Lactic Acid as a platform chemical has broad application in various industries, especially in the production of Poly Lactic Acid (PLA) for biodegradable plastic. Empty fruit bunch (EFB), abundant by product from palm oil mill industry, is one of potential feedstock to be used in the production of lactic acid from lignocellulose biomass. EFB contains high cellulose and hemicellulose about 37– 59.7% w/w and 16–28% w/w, respectively. The aim of this paper is to study the effects of the operating conditions, such as temperature, reaction time, biomass loading, and catalyst concentration on the yield of lactic acid using barium hydroxide as alkaline catalyst. EFB pretreatment with steam explosion was applied to remove lignin content. The results showed that pretreatment reduced the lignin content from 22.66% to 9.69% w/w. Meanwhile, hemicellulose and cellulose increased from 14.40% to 16.40% w/w and 29.37% to 63.57% w/w, respectively. The highest yield of lactic acid was 21.57% C-mol, achieved by using 0.25 M Ba(OH)2 as the catalyst, with 5% w/v biomass loading, temperature 240°C, during 4 h reaction times. The yield was approximately equal to yield of lactic acid (~ 20%) compared with Pb2+ as the catalyst for EFB conversion although the later catalyst produced fewer by products during conversion.

In the current work, date wastes were used for lactic acid (LA) production under thermo-alkaline conditions to overcome some fermentation challenges. Amongst 27 bacterial isolates that produced LA from date juice, isolate D-218 exhibited better growth stability and LA production under various stressed conditions. This isolate was characterized as Bacillus coagulans D-218 using physiological and molecular identification methods. In batch fermentation mode, strain D-218 could not completely utilize 80 g/L of the total sugar and produced only 45.8 ± 2.8 g/L of LA at LA productivity of 0.214 g/L.h and high residual sugar (29.2 g/L) was unutilized in the fermentation media. Different repeated batch fermentations with different initial sugar concentrations and gradual increase in sugar concentrtions were conducted to maximize LA productivity. Starting with 60 g/L of total sugar, four runs of repeated batch fermentations were conducted that enhanced the LA productivity (70%) to 0.49 g/L.h. Another 9 runs were initiated with 40 g/L which further improved LA productivity that reached up to 0.77 g/L.h. Surprisingly, initiating fermentations with 20 g/L of total sugar successfully attained long-term fermentation (18 runs) with high LA yield and productivity without carbon loss of initiated startup sugar of date waste. In this process, strain D-218 could completely consume 80 g/L sugars with high LA production titer (72.9 ± 0.56 g/L), yield (0.92 g/g), and productivity (0.71 g/L.h). This study is the first to exploit date waste in a cost-effective system for high-titer lactic acid production under thermo-alkaline conditions (pH 9.0; 50 °C) and to report repeated batch fermentation for LA production from date wastes.

Lactic acid fermentation from food waste with indigenous microbiota: Effects of pH, temperature and high OLR

Lignocellulosic and algal biomass are promising substrates for lactic acid (LA) production. However, lack of xylose utilization and/or sequential utilization of mixed-sugars (carbon catabolite repression, CCR) from biomass hydrolysates by most microorganisms limits achievable titers, yields, and productivities for economical industry-scale production. This study aimed to design lignocellulose-derived substrates for efficient LA production by a thermophilic, xylose-utilizing, and inhibitor-resistant Bacillus coagulans Azu-10. This strain produced 102.2 g/L of LA from 104 g/L xylose at a yield of 1.0 g/g and productivity of 3.18 g/L/h. The CCR effect and LA production were investigated using different mixtures of glucose (G), cellobiose (C), and/or xylose (X). Strain Azu-10 has efficiently co-utilized GX and CX mixture without CCR; however, total substrate concentration (>75 g/L) was the only limiting factor. The strain completely consumed GX and CX mixture and homoferemnatively produced LA up to 76.9 g/L. On the other hand, fermentation with GC mixture exhibited obvious CCR where both glucose concentration (>25 g/L) and total sugar concentration (>50 g/L) were the limiting factors. A maximum LA production of 50.3 g/L was produced from GC mixture with a yield of 0.93 g/g and productivity of 2.09 g/L/h. Batch fermentation of GCX mixture achieved a maximum LA concentration of 62.7 g/L at LA yield of 0.962 g/g and productivity of 1.3 g/L/h. Fermentation of GX and CX mixture was the best biomass for LA production. Fed-batch fermentation with GX mixture achieved LA production of 83.6 g/L at a yield of 0.895 g/g and productivity of 1.39 g/L/h.

Optimized feeding schemes of simultaneous saccharification and fermentation process for high lactic acid titer from sugarcane bagasse

Dilute sulfuric acid fractionation of Korean food waste for ethanol and lactic acid production by yeast

Lactic acid fermentations were conducted using water hyacinth. It is known that the pretreatment and enzyme hydrolysis process optimize the potential of water hyacinth. Lactic acid produced by using lactic acid bacteria. All cells were grown at <TEX>$37^{\circ}C$</TEX> and initial pH 5.5. Lactic acid production was measured by HPLC. All Lactobacillus strains could produce lactic acid from pretreated water hyacinth. The highest lactic acid was achieved when lactic acid fermentation was carried out by L. delbrueckii for D-form and L. helveticus for L-form lactic acid production. The lactic acid concentration was 10.70 g/L by L. delbrueckii and it converted glucose in the medium to lactic acid, almost perfectly. Lactic acid production became higher when fermentation was carried out at a controlled pH 5.5. Lactic acid yield and productivity were 0.52 g/g and 0.19 g/L/h for L. helveticus, while L. delbrueckii was 0.64 g/g and 0.27 g/L/h. This study showed that water hyacinth medium could be alternative medium which can replace the complex and expensive medium for growing Lactobacillus strains in production of lactic acid.

Production of lactic acid from paper sludge using acid-tolerant, thermophilic Bacillus coagulan strains

Photocatalytic conversion of biomass is considered an effective, clean, and environmentally friendly route to obtain high-valued chemicals and hydrogen. However, the limited conversion efficiency and poor selectivity are still the main bottlenecks for photocatalytic biomass conversion. Herein, we report the highly selective photocatalytic conversion of glucose solution on holo-symmetrically spherical three-dimensionally ordered macroporous TiO₂-CdSe heterojunction photonic crystal structure (s-TCS). The obtained s-TCS photocatalysts show excellent stability and strong light harvesting, uniform mass diffusion and exchange, and efficient photogenerated electrons/holes separation and utilization. The optimized s-TCS-4 photocatalyst displays the highest photocatalytic performance for glucose oxidation and hydrogen production. The glucose conversion, lactic acid selectivity, and yield on s-TCS-4 are about 95.9%, 94.3%, and 96.4%, respectively. The photocatalytic production of lactic acid for s-TCS-4 (18.5 g/L) is 2.3 times higher than the pure spherical TiO₂ photonic crystal without CdSe (s-TiO₂, 8.1 g/L), and the hydrogen production rate of s-TCS-4 is 9.4 times that of s-TiO₂. For the first time, we reveal that the photocatalytic conversion of glucose to lactic acid is a third-order and four-electron-involved reaction. This work could shed some new light on the efficient photocatalysis conversion of biomass to highly value-added products with high selectivity and yield, and simultaneously sustainable hydrogen evolution.

Lactic acid is widely used in the food, chemical, textile, and pharmaceutical industries. New applications of lactic acid for the manufacture of biodegradable polymers have increased the demand for it. Lactic acid can be produced from cheese whey and starchy and lignocellulosic biomass via microbial fermentation with lactic acid bacteria (LAB) or fungi. Pure sugar and cheese whey can be directly fermented by lactic acid bacteria, while liquefaction and enzymatic saccharification of starchy biomass and pretreatment and enzymatic hydrolysis of lignocellulosic biomass are required for lactic acid production from biomass. Amylolytic lactic acid bacteria can direct convert starchy biomass to lactic acid. Lactic acid bacteria and methods used for lactic acid production from different feedstocks are summarized in this paper. Lactic acid productivity of 6.34 and 4.87 g/l·h and yields of 0.98 g/g lactose and 0.97 g/g glucose were obtained from cheese whey and wheat starch, respectively, using cell-recycle repeated batch fermentation by Lactobacillus sp. RKY2. Lactic acid bacteria such as Lactobacillus pentosus, Lactobacillus brevis and Lactococcus lactis can ferment glucose to lactic acid by homolactic fermentation and also effectively convert xylose or arabinose to lactic acid and acetic acid by heterolactic fermentation. The process for lactic acid production from lignocellulosic biomass needs to be improved to increase the lactic acid yield and productivity.

‘Lactobacillus sp. strain TERI-D3’, as microbial cell factory for fermentative production of lactic acid’.

The lespedeza stalks with steam pretreatment were fermented to lactic acid by simultaneous saccharification and fermentation (SSF) in this study. Orthogonal design methodology was used to evaluate the optimum SSF conditions that give maximum lactic acid yield. We have investigated the following relative factors, such as temperature, loading of cellulase, calcium carbonate and concentration of substrate. The optimum operating conditions were found to be temperature of 43 °C, cellulase loading of 30 FPU/g substrate, calcium carbonate of 3 % and substrate of 6 %. Comparisons of different steam pretreated conditions on lactic acid yield from lespedeza stalks were also made. The results showed that lactic acid yields from lespedeza stalks with 4 min pretreatment at pressure of 1.0, 1.25, 1.5 and 2.0 Mpa were 41.8 %, 42.5 %, 50.6 % and 64.0 % of the theoretical, respectively. The lactic acid yield from steam pretreated lespedeza stalks was much higher than that of lespedeza stalks without pretreatment (23.9 %). It can be concluded that the lactic acid yield was remarkably improved by steam pretreatment. The yield of lactic acid from steam pretreated lespedeza stalks was 1.68 times higher than that of untreated ones. Additionally, the lactic acid yield could be further promoted from 64.0 % to 89.4 % by washing pretreated stalks with water, which suggested that water processing is a promising method to remove inhibitors in broth to improve lactic acid yield.

BackgroundEfficient conversion of lignocellulosic biomass to optically pure lactic acid is a key challenge for the economical production of biodegradable poly-lactic acid. A recently isolated strain, Thermoanaerobacterium aotearoense SCUT27, is promising as an efficient lactic acid production bacterium from biomass due to its broad substrate specificity. Additionally, its strictly anaerobic and thermophilic characteristics suppress contamination from other microoragnisms. Herein, we report the significant improvements of concentration and yield in lactic acid production from various lignocellulosic derived sugars, achieved by the carbon flux redirection through homologous recombination in T. aotearoense SCUT27.ResultsT. aotearoense SCUT27 was engineered to block the acetic acid formation pathway to improve the lactic acid production. The genetic manipulation resulted in 1.8 and 2.1 fold increase of the lactic acid yield using 10 g/L of glucose or 10 g/L of xylose as substrate, respectively. The maximum l-lactic acid yield of 0.93 g/g glucose with an optical purity of 99.3% was obtained by the engineered strain, designated as LA1002, from 50 g/L of substrate, which is very close to the theoretical value (1.0 g/g of glucose). In particular, LA1002 produced lactic acid at an unprecedented concentration up to 3.20 g/L using 10 g/L xylan as the single substrate without any pretreatment after 48 h fermentation. The non-sterilized fermentative production of l-lactic acid was also carried out, achieving values of 44.89 g/L and 0.89 g/g mixed sugar for lactic acid concentration and yield, respectively.ConclusionsBlocking acetic acid formation pathway in T. aotearoense SCUT27 increased l-lactic acid production and yield dramatically. To our best knowledge, this is the best performance of fermentation on lactic acid production using xylan as the sole carbon source, considering the final concentration, yield and fermentation time. In addition, it should be mentioned that the performance of non-sterilized simultaneous fermentation from glucose and xylose was very close to that of normal sterilized cultivation. All these results used the mutant strain, LA1002, indicated that it is a new promising candidate for the effective production of optically pure l-lactic acid from lignocellulosic biomass.