Solibacillus daqui sp. nov., isolated from high-temperature Daqu.

A Gram-stain-positive, rod-shaped, endospore-forming, aerobic bacterial strain, designated ZS111008T, was isolated from high-temperature Daqu, a starter for production of Chinese Jiang-flavour Baijiu, and was characterized by polyphasic taxonomy. This novel isolate grew in the presence of 0-5 % (w/v) NaCl, at pH 6.0-9.0 and 25-45 °C; optimum growth was observed with 1 % (w/v) NaCl, at pH 8.0 and 30 °C. A comparative analysis of the 16S rRNA gene sequence (1461 bp) of strain ZS111008T showed highest similarity to Solibacillus silvestris DSM12223T (96.7%), followed by Solibacillus cecembensis PN5T (96.6%) and Solibacillus isronensis AMCK01000046 (96.5%). The DNA G+C content of strain ZS111008T was 37.21 mol%. The respiratory quinone was identified as menaquinone-7 and the major polar lipids were diphosphatidylglycerol, phosphatidylglycerol, phosphatidylethanolamine, phosphatidylserine and one unknown phospholipid. Lys was detected as the diagnostic diamino acid in the cell wall. Based on morphological characteristics, chemotaxonomic characteristics and physiological properties, strain ZS111008T represents a novel species of the genus Solibacillus, for which the name Solibacillus daqui sp. nov. is proposed. The type strain for this proposed species is ZS111008T (=CGMCC 1.19455T=JCM 35214T).

Lentibacillus daqui sp. nov., isolated from high-temperature Daqu, a starter for production of Chinese Jiang-flavour Baijiu.

A Gram-stain-positive, rod-shaped, endospore-forming, aerobic bacterial strain, designated ZS110521T, was isolated from high-temperature Daqu, a starter for production of Chinese Jiang-flavour Baijiu and was characterised by polyphasic taxonomy. This novel isolate grew in the presence of 0-20 % (w/v) NaCl, at pH 6.0-9.0 and 20-50 °C; optimum growth was observed with 8-10 % (w/v) NaCl, at pH 7.0 and 37 °C. A comparative analysis of the 16S rRNA gene sequence (1460 bp) of ZS110521T revealed that it displayed the highest similarity to Lentibacillus populi WD4L-1T (95.5 %), followed by Lentibacillus garicola SL-MJ1T (95.4 %) and Lentibacillus lacisalsi BH260T (95.2 %). ANI and dDDH values between ZS110521T and other strains of species of the genus Lentibacillus were less than 78 and 28 %, respectively. The predominant cellular fatty acids (> 10 %) of ZS110521T were anteiso-C17 : 0 (37.8 %), anteiso-C15 : 0 (28.1 %) and iso-C16 : 0 (15.5 %). The respiratory quinone was identified as menaquinone-7 (MK-7) and the major polar lipids were diphosphatidylglycerol and phosphatidylglycerol. The polyphasic taxonomic data and the results of chemotaxonomic analysis confirmed that ZS110521T represents a novel species, for which the name Lentibacillus daqui sp. nov. is proposed. The type strain of this proposed species is ZS110521T (=CGMCC 1.19456T =JCM 35213T).

Novisyntrophococcus fermenticellae gen. nov., sp. nov., isolated from an anaerobic fermentation cellar of Chinese strong-flavour baijiu.

A Gram-stain-negative, coccus-shaped, obligately anaerobic, non-motile and non-spore-forming bacterium, designated strain JN500902T, was isolated from the mud in a fermentation cellar used continuously over 30 years for Chinese strong-flavour baijiu production. Colonies were white, circular, convex and smooth-edged. Growth was observed at 20-40 °C (optimum, 37 °C), at pH 5.0-10 (optimum, pH 7.5), with 0-2 % (w/v) NaCl and with 0-4 % (v/v) ethanol. The Biolog assay demonstrated positive reactions of strain JN500902T in the metabolism of l-fucose and pyruvate. The predominant cellular fatty acids (>10 %) consisted of C16 : 0 and C14 : 0. The major end metabolites of strain JN500902T were acetic acid and ethanol when incubated anaerobically in liquid reinforced clostridial medium. Acetate was the major organic acid end product. The complete genome size of strain JN500902T was 3 420 321 bp with 3327 identified genes. The G+C content was 43.5 mol%. Phylogenetic analysis based on 16S rRNA gene sequences affiliated strain JN500902T with the family Lachnospiraceae, having low sequence similarity (92.8 %) to the nearest type strain, Syntrophococcus sucromutans DSM 3224T and forming a clearly distinct branch. Core genome phylogenetic analysis of the isolate and 134 strains belonging to the family Lachnospiraceae also revealed that strain JN500902T was well-separated from other genera of this family as a monophyletic clade. The average nucleotide identity and amino acid identity values between strain JN500902T and 134 Lachnospiraceae strains were less than 74 and 65 %, respectively. Considering its polyphasic characteristics, strain JN500902T represents a novel genus and species within the family Lachnospiraceae, for which the name Novisyntrophococcus fermenticellae gen. nov., sp. nov. is proposed. The type strain is JN500902T (=CICC 24502T=JCM 33939T).

Silage Fermentation on Sweet Sorghum Whole Plant for Fen-Flavor Baijiu.

The technology for producing bioethanol from sweet sorghum stalks by solid-state fermentation has developed rapidly in recent years, and has many similarities with traditional Chinese liquor production. However, the product from sweet sorghum stalks was lacking in volatile flavors, and the level of harmful contents were uncertain, therefore it could not be sold as liquor. In this study, the protein, fat, and tannin in the clusters and leaves of sweet sorghum were utilized to increase the content of flavor compounds in the ethanol product through the anaerobic fermentation of Saccharomyces cerevisiae. Meanwhile, the silage fermentation method was used to extend the preservation time of the raw materials and to further enhance the flavors of Fen-flavor liquor, with ethyl acetate as the characteristic flavor. The effects of different feedstock groups on ethyl acetate, ethyl lactate, methanol, acetaldehyde, acetal, fusel oil, total acid, and total ester were evaluated by analyzing the chemical composition of different parts of sweet sorghum and determined by gas chromatograph. The effect of different fermentation periods on the volatile flavor of sweet sorghum Baijiu was evaluated. The yield of the characteristic volatile flavor was increased by the extension of the fermentation time. Sweet sorghum Baijiu with a high ester content can be used as a flavoring liquor, blended with liquor with a shorter fermentation period to prepare the finished Fen-flavor Baijiu, conforming to the Chinese national standard for sale.

Identification of water-soluble peptides in distilled spent grain and its angiotensin converting enzyme (ACE) inhibitory activity based on UPLC-Q-TOF-MS and proteomics analysis.

Distilled spent grain (DSG) is the biggest by-product in baijiu (Chinese liquor) production, releasing approximately 23.44 million tons every year. Aiming at comprehensive identification of more bioactive peptides, in this work, the new bioassay-guided proteomics and Biolynx peptide sequencer based on ultra-performance liquid chromatography quadrupole time-of-flight mass spectrometry (UPLC-Q-TOF-MS) were developed. Moreover, 22 peptides with angiotensin converting enzyme (ACE) inhibitory activities were identified. Seven peptides were successfully quantified using electrospray ionization with triple-quadrupole mass spectrometry (ESI-QQQ-MS) in the multiple reaction monitoring (MRM). Of these identified peptides, Pro-Arg was the most abundant (92.14 µg g-1 dry weight (DW)) and acted as a competitive inhibitor of ACE by molecular docking. Therefore, peptides from DSG can be considered as promising candidates for ACE inhibition; in addition, the new strategy for peptide sequencing can be extended to any food matrices containing peptide mixture or protein hydrolysate.

Determination and comparison of flavor (retronasal) threshold values of 19 flavor compounds in Baijiu.

Nineteen compounds, including ten esters, six acids, and three alcohols, were characterized and considered as significant tastants and aromas in Baijiu (Chinese Liquor). The flavor (retronasal) threshold values (FTVs) of these 19 compounds were determined by the 10 samples test method in hydroalcoholic solutions (46% v/v in ethanol). The FTVs of the compounds were calculated based on the best estimate threshold method. All the FTVs determined by the professional Chinese Baijiu tasters were lower than those by the nonprofessional tasters. For instance, the detection (2.31 mg/kg) and recognition (11.74 mg/kg) values of ethyl hexanoate determined by the nonprofessional group were higher than the respectively corresponding values 0.44 and 3.80 mg/kg determined by the professional group. All of the odor activity values (OAVs) of ethyl valerate (OAV: 1176.00 to 2321.17), ethyl octanoate (OAV: 6841.20 to 7851.60), and 1-butanol (OAV: 26.78 to 39.72) in Gujinggong Baijiu were more than 10-fold larger than their dose-over-threshold values (DoTs), for which the DoTs of ethyl valerate, ethyl octanoate, and 1-butanol were 92.84 to 183.25, 180.03 to 206.62, 1.18 to 1.75, respectively. On the contrary, the OAVs of ethyl heptanoate (OAV: 3.60 to 5.70) and isoamyl alcohol (OAV: 1.18 to 1.57) were lower than their corresponding DoTs at 152.62 to 241.63 and 12.26 to 16.41. The results demonstrated that it is necessary to consider and compare their DoTs and OAVs simultaneously on evaluating the contribution of flavor compounds in Baijiu. PRACTICAL APPLICATION: Sensory evaluation of threshold values of various flavor compounds could be significantly affected by their existing matrix. Most of the published results of the flavor threshold value of compounds were determined from the matrix such as beer, whiskey, red wine, rather than Chinese Baijiu. The results of this work not only could provide valuable information for flavor studies of Chinese Baijiu but also give useful information for the Baijiu industry to quality control.

In Vitro Production and Identification of Angiotensin Converting Enzyme (ACE) Inhibitory Peptides Derived from Distilled Spent Grain Prolamin Isolate.

Distilled spent grain (DSG), the biggest by-product of the Chinese liquor industry, is rich in protein (167.8 g/kg DSG dry weight (DW)). Accounting for 60% of the total protein, prolamins are isolated from dried DSG (DDSG). In this study, angiotensin-converting enzyme (ACE) inhibitory peptides were screened from the prolamin hydrolysates of DDSG using two independent active-directed separations, ultrafiltration and reversed phase high performance liquid chromatography (RP-HPLC) coupled with ACE inhibitory activity evaluation. Six novel ACE inhibitory peptides, AVQ, YPQ, NQL, AYLQ, VLPVLS, and VLPSLN, were successfully identified and quantified from the active RP-HPLC fractions. AVQ and YPQ exhibited the highest activity, having the concentration inducing 50% inhibition (IC50) values for ACE of 181.0 and 220.0 µM, respectively. It was observed that VLPVLS was the most abundant peptide (16.96 mg/g DW) in prolamins. The results indicated that prolamin hydrolysates from DDSG could be served as a source of ACE inhibitory peptides.