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If a pure culture of ''Lactobacillus'' bacteria is used it is ideal but not necessary to fill the head space of the fermenter with CO2 gas (some brewers have reported that this will help reduce sulfur in the finished beer). Keeping positive pressure in the kettle with CO2 will help prevent contaminates that create [[Butyric Acid|butyric acid]] and other off-flavors from entering the kettle due to negative pressure, and is often the approach that commercial brewers take <ref>Personal correspondence with Steph Cope of CraftHaus Brewing Co. 02/06/2016.</ref>. The kettle should be held at the desired temperature for 24-72 hours (in some cases longer, but no longer than 5 days). Depending on the strain of ''Lactobacillus'', and the desired sour level, the time of incubation is ultimately a variable that is up to the brewer (see the ''[[Lactobacillus]]'' page for suggested temperatures and times for specific strains). The kettle lid should be firmly in place and optionally sealed with plastic wrap so that other microorganisms do not get in. Potential for the formation of [[Butyric Acid]] and [[Isovaleric Acid]] when using only a pure culture is extremely slight to none assuming contamination does not occur. Temperature shifts during kettle souring are not a concern as long as the temperature does not get too hot or too cold for the specific ''Lactobacillus'' culture. Some species, such as ''L. plantarum'', create acidity at room temperature, and so some brewers will pitch this strain at around 90-100°F, and then let the temperature fall to room temperature during souring. Other species might not perform as well at colder temperatures, and so maintaining a fairly consistent hot temperature is desirable. If the temperature is allowed to fall, take precautions on not allowing any dust to get sucked into the fermenter since temperature decreases will create a vacuum inside the fermenter (flushing with CO2 is a good way to prevent a vacuum).
In order to address the challenges of creating a sealed environment in a commercial boil kettle (which is often left open to the environment through the boil stack), some commercial brewers have found it beneficial to rack the wort from the boil kettle to a mashtun, brite tank, fermenter (often dedicated), or another vessel that seals and purges well to do the souring in. The mashtun or other vessel is first cleaned, sanitized, and often purged with CO2. Once the wort is in the vessel and at the correct temperature for the given culture of ''Lactobacillus'', the ''Lactobacillus'' is pitched into the vessel and the vessel is sealed off. If the vessel is sealed and air tight, temperature shifts from cooling won't suck air into the vessel, and a potentially more sanitary souring fermentation will occur. Using a fermenter or brite tank (if there are multiple) has the advantage of not occupying a bottleneck vessel such as the mashtun or boil kettle, but has more risk if the fermenter is not dedicated to kettle souring. Dedicating removable soft parts like gaskets and hoses will minimize the risk of infection originating from the fermenter or brite tank if it is not dedicated to souring <ref>[https://www.facebook.com/groups/MilkTheFunk/permalink/1778027798892093/ Various professional brewers. Milk The Funk Facebook post about souring in a mashtun, brite tank, or other vessel for commercial brewers. 08/02/2017.]</ref>.
Once the level of acidity is reached (this can be tested with a reliable [[PH_Meter|pH meter]], or in the case of using a pure culture can safely be taste tested), the wort is brought to a boil. The wort may be boiled normally in the case of any style of beer that requires a longer boiling process, or it may be boiled for no more than a minute or two in the case of making a [[Berliner Weissbier]]. Technically speaking, the wort doesn't need to be boiled at all (this is called [http://www.garshol.priv.no/blog/331.html Raw Ale]). Heat pasteurization at 140°F (60°C) for 15 minutes should kill the ''Lactobacillus'' culture being used to sour the wort <ref name="pasteurization" />.